Modulators of TNF-α signaling

ABSTRACT

The present invention provides compounds which are modulators of TNF-α signaling and methods of use thereof for treating a patient having a TNF-α mediated condition. The compounds can be represented by the following structural formulas:

RELATED APPLICATIONS

This application is a continuation application of U.S. Ser. No.13/946,389 filed Jul. 19, 2013, which is a divisional application ofU.S. Ser. No. 11/292,325, filed Dec. 1, 2005, now U.S. Pat. No.8,518,999, which is a continuation application of U.S. Ser. No.10/797,244, filed Mar. 10, 2004, now U.S. Pat. No. 7,034,031, which is adivisional application of U.S. Ser. No. 09/852,965, filed May 10, 2001,now U.S. Pat. No. 6,969,728, which claims the benefit of U.S.Provisional Application No. 60/203,784, filed May 12, 2000, and U.S.Provisional Application No. 60/205,213, filed May 18, 2000. The entireteachings of these applications are incorporated herein by reference.

BACKGROUND OF THE INVENTION

The cytokine tumor necrosis factor α (“TNF-α”) has a broad spectrum ofbiological activities. TNF-α is produced by activated macrophages and avariety of other cells, including antigen-stimulated T cells, activatedmast cells and activated natural killer cells. TNF-α is initiallyproduced as a transmembrane protein of about 25 kD. A 17 kD fragment ofthis membrane protein is proteolytically cleaved from the cell membraneand circulates as a 51 kD homotrimer. TNF-α mediated processes proceedvia the interaction of this trimeric protein with a receptor protein atthe surface of a target cell.

TNF-α plays an important role in coordinating the body's response toinfection, and serves as an important mediator of inflammation. Forexample, TNF-α signaling has been implicated in the induction of feverand the production of interferon-α by T cells. TNF-α induces increasedbinding of leukocytes to endothelial cells, resulting in accumulation ofleukocytes at sites of infection. TNF-α signaling has also beenimplicated in inducing the production of interleukin-1 andprostaglandins by macrophages, and is involved in the breakdown of theextracellular matrix, inducing collagenase in synoviocytes, and in boneresorption via osteoclast activation.

TNF-α has certain effects on the growth and metastatic potential oftumors. For example, certain human tumor cell lines are sensitive toTNF-α in vitro and TNF-α activation may precede killing of tumor cellsby macrophages.

High levels of TNF-α are generally associated with chronic immune orinflammatory diseases, and are considered a cause of neural and cellulardegeneration. At lower levels, however, TNF-α plays an important role inthe cell life cycle, cellular response to foreign attack, andmaintenance of homeostasis. For this reason, it will be appreciated thatthe purpose of this invention is not the complete and absoluteinhibition of TNF-α, but rather the modulation of the cellular responseto TNF-α levels and the treatment of TNF-α mediated conditions, therebypermitting an effective treatment for the chronic immune andinflammatory responses that occur when excess TNF-α is produced.

The production of TNF-α has been implicated in a variety of diseasestates including but not limited to the following: septic shock;endotoxic shock; cachexia syndromes associated with bacterialinfections, such as tuberculosis and meningitis; viral infections, suchas AIDS; parasitic infections, such as malaria; neoplastic disease;autoimmune disease, including some forms of arthritis, especiallyrheumatoid and degenerative forms; and adverse effects associated withtreatment for the prevention of graft rejection. Thus, there is a needfor agents which can interrupt or modulate the TNF-α signaling process.

SUMMARY OF THE INVENTION

The present invention provides compounds which are modulators of TNF-αsignaling and methods of use thereof for treating a patient having aTNF-α mediated condition.

In one embodiment, the invention provides compounds of Formula (I),

In Formula (I), R₁ is H or NH₂; R₂ and R₃ are each, independently, —H,—OH, a substituted or unsubstituted alkyl, or a substituted orunsubstituted alkoxy; R₄ is, —H or a substituted or unsubstituted alkyl;X is O, S, CH₂ or SO₂; V, W and Z are each, independently, N or CH; Y issubstituted and unsubstituted phenyl or a substituted and unsubstitutedheterocyclyl; and n is 0, 1 or 2.

In another embodiment, the invention provides compounds of Formula II,

where R₅ is substituted or unsubstituted aralkyl, a substituted orunsubstituted cycloalkyl or a substituted or unsubstitutedcycloalkylalkyl; R₆ is —H or —NR₁₃R₁₄; R₇ is substituted orunsubstituted phenyl; and R₁₃ and R₁₄ are each, independently, —H, asubstituted or unsubstituted alkyl, a substituted or unsubstitutedcycloalkyl, a substituted or unsubstituted aryl, a substituted orunsubstituted aralkyl or R₁₃ and R₁₄ together with the nitrogen to whichthey are attached is a heterocycloalkyl.

The present invention further relates to compounds of Formula III,

where R₈ and R₁₂ are each, independently, —H, a substituted orunsubstituted alkyl, a substituted or unsubstituted aryl, a substitutedor unsubstituted aralkyl or a substituted or unsubstitutedheteroaralkyl; R₉ is —H, a substituted or unsubstituted aryl, asubstituted or unsubstituted aralkyl, a substituted or unsubstitutedheteroaryl or a substituted or unsubstituted heteroaralkyl; R₁₀ issubstituted or unsubstituted alkyl, a substituted or unsubstituted aryl,a substituted or unsubstituted heteroaralkyl or a substituted orunsubstituted heterocycloalkylalkyl; and R₁₁ is substituted orunsubstituted alkyl, a substituted or unsubstituted aryl, a substitutedor unsubstituted aralkyl, a substituted or unsubstitutedcycloalkylalkyl, a substituted or unsubstituted heteroaryl, asubstituted or unsubstituted heteroaralkyl, a substituted orunsubstituted benzophenonyl or a substituted or unsubstitutedcycloalkylalkyl.

In yet another embodiment, the present invention relates to a method oftreating a TNF-α mediated condition in a patient. The method comprisesthe step of administering to the patient a therapeutically effectiveamount of at least one compound of Formula I, Formula II or Formula III,as defined above.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to compounds which are antagonists ofTNF-α signaling and, therefore, are effective agents for the treatmentof TNF-α mediated medical conditions, disorders and diseases, such aschronic inflammation, tissue breakdown and cancer.

For the purposes of the present invention, the language “alkyl” isintended to include a straight chain or branched saturated hydrocarbylgroup. Preferred alkyl groups include C₁-C₁₂-alkyl groups, while morepreferred alkyl groups include C₁-C₆-alkyl groups. The language“cycloalkyl” is intended to include a mono-, bi- or polycyclic alkylgroup. Preferred cycloalkyl groups include monocyclic C₃-C₈-cycloalkylgroups. The language “alkoxy” is intended to include an alkyl-O-group ora cycloalkyl-O-group, where the preferred alkyl and cycloalkyl groupsare those given above. The language “aromatic ether” is intended toinclude an —O-aryl or —O-heteroaryl. The language “alkenyl” is intendedto include a straight chain or branched hydrocarbyl group which includesone or more double bonds. Preferred alkenyl groups includeC₂-C₁₂-alkenyl groups. The language “cycloalkenyl” is intended toinclude a cyclic hydrocarbyl group which includes one or more doublebonds but is not aromatic. Preferred cycloalkenyl groups includeC₅-C₈-cycloalkenyl groups.

The language “aryl” is intended to include an aromatic carbocyclicgroup, such as a phenyl group, a naphthyl group or a phenyl or naphthylgroup which is fused with a five or six-membered saturated, partiallyunsaturated or aromatic carbocyclic ring.

The language “heterocycle” and “heterocyclic group” is intended toinclude a saturated, aromatic or partially unsaturated ring system whichincludes at least one heteroatom, such as one or more oxygen, nitrogenor sulfur atoms or a combination thereof.

The language “heterocycloalkyl” is intended to include saturatedheterocyclic groups, such as piperidyl, pyrrolidyl, piperazyl,tetrahydrofuranyl and morpholyl.

The language “heteroaryl” is intended to include an aromaticheterocyclic group. Suitable heteroaryl groups include, but are notlimited to, pyridyl, pyrimidyl, quinolyl, isoquinolyl, pyrrolyl,quinoxalyl, imidazolyl, oxazolyl, isoxazolyl, pyrazolyl, thienyl,furanyl, pyrazolyl, thiadiazolyl, oxadiazolyl, indazolyl, thiazolyl,isothiazolyl, and tetrazolyl. Heteroaryl groups also include ringsystems in which a carbocyclic aromatic ring, carbocyclic non-aromaticring or heteroaryl ring is fused to one or more other heteroaryl rings,e.g., benzo(b)thienyl, benzimidazolyl, benzoxazolyl, benzothiazolyl,benzothiadiazolyl, benzoxadiazolyl, indolyl, tetrahydroindolyl,azaindolyl, indazolyl, quinolinyl, imidazopyridyl, puryl,pyrrolo[2,3-d]pyrimidyl, pyrazolo[3,4-d]pyrimidyl.

The language “aralkyl” is intended to include an alkyl group which issubstituted by one or more aryl groups. A substituted aralkyl can have asubstitutent on the aryl or on the alkyl portion of the aralkyl.Preferred aralkyl groups include benzyl, diphenylmethyl and 2-phenethylgroups. The language “heteroaralkyl” is intended to include an alkylgroup which is substituted by a heteroaryl group or by a heteroarylgroup and one or more aryl groups. A substituted heteroaralkyl can havea substituent on the a heteroaryl or on the alkyl portion of theheteroaralkyl. Preferably, a heteroaryl group is an alkyl groupsubstituted by a heteroaryl group.

The language “cycloalkylalkyl” is intended to include an alkyl groupsubstituted with a cycloalkyl group.

The language “heterocycloalkylalkyl” is intended to include an alkylgroup substituted with a heterocycloalkyl group.

Alkyl, cycloalkyl, alkenyl, cycloalkenyl, the alkyl portion of anaralkyl, the alkyl portion of a heteroarlkyl, cycloalkylalkyl, andalkoxy groups can be substituted or unsubstituted. Substituted groups ofthis type can include one or more substituents independently selectedfrom halo, such as fluoro, chloro, bromo and iodo; alkyl, such asC₁-C₆-alkyl; nitro; hydroxyl; —NR₁₃R₁₄, wherein R₁₃ and R₁₄ are definedas above; —C(O)R₁₅, wherein R₁₅ is —H, an alkyl, an aryl or an aralkyl;cyano; aryl groups; cycloalkyl groups and heterocyclic groups, such asheteroaryl groups.

Aryl, heterocyclic, such as heteroaryl, aromatic ethers, the aromaticportion of an aralkyl, the aromatic portion of heteroaralkyl,heterocycloalkylalkyl and benzophenone groups can be substituted orunsubstituted. Suitable substituents include one or more substituentsindependently selected from halo, such as fluoro, chloro, bromo or iodo;alkyl, preferably C₁-C₃-alkyl; a halogenated alkyl, preferably ahalogenated C₁-C₃-alkyl; alkoxy, preferably C₁-C₃-alkoxy; aromaticether; alkyl substituted with an aromatic ether; a hydroxy substitutedalkyl; alkoxy substituted aromatic ether; —S-(alkyl); cyano; azide;nitro; —C(O)R₁₅; —NR₁₃R₁₄; —C(O)NR₁₃R₁₄; —C(O)OR₁₆, wherein R₁₆ is —H,an alkyl, an aryl or an aralkyl; benzyl;4-(4-benzylpiperazin-1-yl)methyl;4-4-(2-fluorophenyl)piperazin-1-yl)methyl; a halogenated aryl;methylenedioxo; an aryl; a heteroaralkyl; a heterocycloalkylalkyl; and aheterocyclic, such as a heteroaryl group.

In one embodiment, the present invention provides compounds of FormulaI,

In Formula (I), R₁ is H or NH₂; R₂ and R₃ are each, independently, —H,—OH, a substituted or unsubstituted alkyl or a substituted orunsubstituted alkoxy; R₄ is, —H or a substituted or unsubstituted alkyl;X is O, S, CH₂ or SO₂; V, W and Z are each, independently, N or CH; Y issubstituted and unsubstituted phenyl or a substituted and unsubstitutedheterocyclyl; and n is 0, 1 or 2.

In one embodiment, Y is a phenyl group which has one or moresubstituents independently selected from the group consisting ofhalogen, linear or branched C₁-C₄-alkoxy, trifluoromethoxy,dioxymethylene, hydroxyalkyl, trifluoromethyl, HC(O)—, linear orbranched C₁-C₄-alkyl, heterocyclyl and substituted or unsubstitutedheterocycloalkylalkyl. Preferred substituents for Y include fluoro,chloro, methoxy, morpholyl, N-morpholinomethyl, tetrahydroisoquinolyl,tetrahydroisoquinolinomethyl, 4-(4-benzyl-piperazin-1-yl)methyl,4-(4-(2-fluoro-phenyl)piperazin-1-yl)methyl and isopropyl. Y can also bea heterocyclyl group, e.g., pyridyl, furyl or pyrrolidyl.

Alternatively in Formula (I), R₁-R₄, V, W, X, Z and n are as describedabove, and Y is represented by the following structural formula:

wherein R₅₀ and R₅₁ are independently an alkyl group, a substitutedalkyl group, an aryl group a substituted aryl group, or, taken togetherwith the nitrogen atom to which they are bonded, are a substitutedheterocycloalkyl, an unsubstituted heterocycloalkyl, a substitutedheteroaryl group or an unsubstituted heteroaryl group. Preferably, R₅₀and R₅₁ are, taken together with the nitrogen atom to which they arebonded, an N-substituted piperazyl group, wherein the N-substituent isan aryl group, a substituted aryl group, a —CH₂-aryl group or a—CH₂-(substituted aryl group), and preferably phenyl, substitutedphenyl, benzyl or substituted phenyl. In a preferred embodiment, R₁ andR₄ are —H, R₂-R₃ are methyl, V and Z are each —CH—, W is —N—, X is —O—and n is 0. Suitable substituents for a heterocycloalkyl or heteroarylgroup formed by R₅₀ and R₅₁ taken together with the nitrogen atom towhich they are bonded are as described below for substitutedheterocycles.

In another embodiment, the invention provides compounds of Formula II,

where R₅ is substituted or unsubstituted aralkyl, substituted orunsubstituted cycloalkyl or substituted or unsubstitutedcycloalkylalkyl; R₆ is —H or —NR₁₃R₁₄; R₇ is substituted orunsubstituted phenyl; and R₁₃ and R₁₄ are each, independently, —H, asubstituted or unsubstituted alkyl, a substituted or unsubstitutedcycloalkyl, a substituted or unsubstituted aryl, a substituted orunsubstituted aralkyl or R₁₃ and R₁₄ together with the nitrogen to whichthey are attached is a heterocycloalkyl.

In one embodiment, R₅ is substituted or unsubstituted benzyl. Suitablesubstituents on the benzyl group include halogen atoms and linear andbranched C₁-C₄-alkoxy. Preferably, R₅ is unsubstituted benzyl or benzylhaving one or more substituents independently selected from chloro andmethoxy. In another embodiment, R₅ is C₃-C₈-cycloalkyl,C₃-C₈-cycloalkyl-C₁-C₄-alkyl or substituted or unsubstitutedphenyl-C₂-C₄-alkyl. For example, R₅ can be 2-phenethyl, cyclohexyl orcyclopentylethyl.

R₇ is, preferably, phenyl having one or more of the followingindependently selected substituents: halogen, linear C₁-C₆-alkyl,branched C₁-C₆-alkyl, cyclic C₃-C₆-alkyl or trifluoromethyl. Morepreferably, R₇ is phenyl having one or more of the followingindependently selected substituents: fluoro, chloro, and linearC₁-C₄-alkyl or branched C₁-C₄-alkyl.

The present invention further relates to compounds of Formula III

where R₈ and R₁₂ are each, independently, —H, a substituted orunsubstituted alkyl, a substituted or unsubstituted aryl, substituted orunsubstituted aralkyl or a substituted or unsubstituted heteroaralkyl;R₉ is —H, a substituted or unsubstituted aryl, a substituted orunsubstituted aralkyl, a substituted or unsubstituted heteroaryl or asubstituted or unsubstituted heteroaralkyl; R₁₀ is substituted orunsubstituted alkyl, a substituted or unsubstituted aryl, a substitutedor unsubstituted heteroaralkyl or a substituted or unsubstitutedheterocycloalkylalkyl; and R₁₁ is substituted or unsubstituted alkyl, asubstituted or unsubstituted aryl, a substituted or unsubstitutedaralkyl, a substituted or unsubstituted cycloalkylalkyl, a substitutedor unsubstituted heteroaryl, a substituted or unsubstitutedheteroaralkyl, a substituted or unsubstituted benzophenone or asubstituted or unsubstituted cycloalkylalkyl.

In a preferred embodiment, one of R₈ and R₁₂ is —H and the other issubstituted or unsubstituted phenyl, phenyl-C₁-C₄-alkyl,diphenyl-C₁-C₄-alkyl, linear C₁-C₁₂-alkyl, branched C₁-C₁₂-alkyl, cyclicC₃-C₁₂-alkyl or dicycloalkyl-C₁-C₄-alkyl. Examples of suitablesubstituents for the phenyl group(s) of R₈ or R₁₂ include one or more ofthe following independently selected groups: alkoxy, such asC₁-C₄-alkoxy, preferably methoxy; alkyl, such as C₁-C₄-alkyl, preferablymethyl and ethyl; and cyano. Suitable identities for R₈ or R₁₂ include,but are not limited to, 2,2-diphenylethyl, 2-(4-ethylphenyl)ethyl,benzyl, diphenylmethyl, 1,2-diphenylethyl, 3,3-diphenylpropyl,3,4,5-trimethoxybenzyl, 2,4,4-trimethylisopentyl,2-(4-methoxyphenyl)ethyl, 2-cyclopentyl-2-phenylethyl, or2-phenyl-2-pyridylethyl.

R₉ is, preferably, substituted or unsubstituted phenyl, a substituted orunsubstituted phenyl-C₁-C₄-alkyl, diphenyl-C₁-C₄-alkyl, phenylfuranyl orheteroaryl-C₁-C₄-alkyl. Suitable phenyl substituents for a substitutedphenyl or a substituted phenyl-C₁-C₄-alkyl include one or more of thefollowing independently selected groups: cyano; alkyl, such asC₁-C₄-alkyl, preferably methyl; alkoxy, such as C₁-C₄-alkoxy, preferablymethoxy; C₁-C₄-alkyl-S—; a halogen, preferably chloro or fluoro; ahalogenated C₁-C₄-alkyl, preferably trifluoromethyl; and phenoxy. Aphenoxy substituent can also be substituted with an alkyl or alkoxygroup as described above. Suitable identities for R₉ include, but arenot limited to, phenyl, 2-cyanophenyl, 3-cyanophenyl, 4-cyanophenyl,diphenylmethyl, pyrazolylmethyl, 2,4-dimethylphenyl, 2-methylphenyl,3-methylphenyl, 4-methylphenyl, 2-methyl-4-methoxyphenyl,3-methyl-4-methoxyphenyl, 4-methylthiophenyl, 3-chlorophenyl,3-trifluoromethylphenyl, benzyl, 2-trifluoromethylbenzyl,3-trifluoromethylbenzyl, 2-chlorobenzyl, 3-chlorobenzyl, 4-chlorobenzyl,2-methoxybenzyl, 3-methoxybenzyl, 4-methoxybenzyl, 2-fluorobenzyl,3-fluorobenzyl, 4-fluorobenzyl, 3-azidylphenyl,3-(4-methoxyphenoxyl)phenyl, or 5-phenylfuran-2-yl.

In one embodiment, R₁₀ is substituted or unsubstituted phenyl, alkylsubstituted with a heteroaryl group, alkyl substituted with aheterocycloalkyl group, or an alkyl substituted with —NR₁₃R₁₄,preferably N,N-dialkylamine. Suitable identities for R₁₀ include, butare not limited to, 2-(imidazol-4-yl)ethyl, 3-(imidazol-4-yl)propyl,3-(imidazol-1-yl)propyl 2-(3-methylimidazol-4-yl)ethyl,2-(morpholin-4-yl)ethyl, 2-(4-pyrazolyl)ethyl, 4-pyrazolylmethyl,2-N,N-dimethylaminoethyl, 3-N,N-dimethylaminopropyl, and2-(aminocarbonyl)phenyl.

R₁₁ is, preferably, a linear or branched C₁-C₄-alkyl, substituted orunsubstituted phenyl, substituted or unsubstituted benzophenonyl,pyrazolyl, aminopyrazolyl, substituted or unsubstitutedindolyl-C₁-C₄-alkyl, thiophenyl, quinoxaline, substituted orunsubstituted phenyl-C₁-C₄-alkyl, pyridylcarbonylphenyl,phenylcarbonyl-C₁-C₄-alkyl, naphthyl, naphthyl-C₁-C₄-alkyl,diphenyl-C₁-C₄-alkyl, C₅-C₈-cycloalkyl-C₁-C₄-alkyl,C₁-C₄-alkylcarbonyl-C₁-C₄-alkyl, fluorenyl, pryrrolyl, N-methylpyrrolyl,or pyridyl. Suitable substituents on the phenyl ring include halogens,preferably fluoro; furanyl; thiophenyl; phenyl; benzyl; phenoxy; alkyl,such as C₁-C₄-alkyl, preferably methyl; phenoxyalkyl;C₁-C₄-alkylcarbonyl; —C(O)-benzyl; and alkoxy, such as C₁-C₄-alkoxy,preferably methoxy. Suitable substituents on the benzophenonyl ringsystem include alkoxy, such as C₁-C₄-alkoxy, preferably methoxy;halogens, preferably chloro; and a C₁-C₄-alkyl group. Suitablesubstituents on the indole ring of a indolyl-C₁-C₄-alkyl includehalogens, preferably bromo. Suitable substituents on the C₁-C₄-alkyl ofa phenyl-C₁-C₄-alkyl include hydroxyl groups. Suitable substituents onthe C₁-C₄-alkyl of a indolyl-C₁-C₄-alkyl include hydroxyl groups.Suitable identities of R₁₁ include, but are not limited to,benzophenon-2-yl, 4′-methoxybenzophenon-2-yl, 4′-chlorobenzophenon-2-yl,2-(furan-2-yl)phenyl, 2-(thiophen-2-yl)phenyl, 2-benzylphenyl,2-pyridylcarbonylphenyl, 2-(phenoxymethyl)phenyl,2-(t-butylcarbonyl)phenyl, 2,2-diphenylethyl, 1-fluorenyl,(naphth-2-yl)methyl, naphth-1-yl, 3-(phenylcarbonyl)propyl,4-phenylbutyl, 4-butylphenyl, 2-(4-chlorophenylcarbonyl)phenyl,3-methoxyphenyl, N-methylpyrrol-2-yl, 2,3-dimethoxyphenyl,3-butyl-2-pyridyl, 2-naphthylmethyl, 2-cyclohexylethyl, 3-methoxyphenyl,N-methyl-2-pyrrolyl, 2-cyclopentylethyl, 3-oxobutyl, 2-benzopyrazyl,quinoxalin-2-yl, 3-idolyl, (2-methylindol-3-yl)methyl,3-(indol-3-yl)propyl, (indol-3-yl)methyl, (5-bromoindol-3-yl)methyl,3-chlorophenyl, 3-aminopyrazol-4-yl, 2-(indol-3-yl)-1-hydroxyethyl,3-fluorophenyl, 1-phenyl-1-hydroxymethyl, 2-phenylphenyl,2-phenoxyphenyl, thiophen-2-yl, and isopropyl.

Certain compounds of formula I, II or III may contain one or more chiralcentres, and exist in different optically active forms. When compoundsof formula I contain one chiral centre, the compounds exist in twoenantiomeric forms and the present invention includes both enantiomersand mixtures of enantiomers. The enantiomers may be resolved by methodsknown to those skilled in the art, for example by formation ofdiastereoisomeric salts which may be separated, for example, bycrystallization; formation of diastereoisomeric derivatives or complexeswhich may be separated, for example, by crystallization, gas-liquid orliquid chromatography; selective reaction of one enantiomer with anenantiomer-specific reagent, for example enzymatic esterification; orgas-liquid or liquid chromatography in a chiral environment, for exampleon a chiral support, for example silica with a bound chiral ligand or inthe presence of a chiral solvent. It will be appreciated that where thedesired enantiomer is converted into another chemical entity by one ofthe separation procedures described above, a further step is required toliberate the desired enantiomeric form. Alternatively, specificenantiomers may be synthesized by asymmetric synthesis using opticallyactive reagents, substrates, catalysts or solvents, or by converting oneenantiomer into the other by asymmetric transformation.

When a compound of formula I, II or III contains more than one chiralcentre it may exist in diastereoisomeric forms. The diastereoisomericpairs may be separated by methods known to those skilled in the art, forexample chromatography or crystallization and the individual enantiomerswithin each pair may be separated as described above. The presentinvention includes each diastereoisomer of compounds of formula I, II orIII and mixtures where the diastereoisomers are unresolved.

Certain compounds of formula I, II, or III may exist in differenttautomeric forms or as different geometric isomers, and the presentinvention includes each tautomer and/or geometric isomer of compounds offormula I, II or III and tautomeric mixtures thereof.

Certain compounds of formula I, II or III may exist in different stableconformational forms which may be separable. Torsional asymmetry due torestricted rotation about an asymmetric single bond, for example becauseof steric hindrance or ring strain, may permit separation of differentconformers. The present invention includes each conformational isomer ofcompounds of formula I, II or III and mixtures of conformational isomersthereof.

Certain compounds of formula I, II or III may exist in zwitterionic formand the present invention includes each zwitterionic form of compoundsof formula I, II or III and mixtures thereof.

The present invention further relates to pharmaceutically acceptablesalts of the compounds of Formulas I, II and III. The phrase a“pharmaceutically acceptable salt” is intended to include a salt whichretains the biological effectiveness and properties of the free base andwhich can be obtained by reaction with an inorganic or organic acid,such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,phosphoric acid, organic sulfonic acid, organic carboxylic acid, organicphosphoric acid, for example, methanesulfonic acid, ethanesulfonic acid,p-toluenesulfonic acid, salicylic acid, lactic acid, tartaric acid andthe like.

Compounds of Formulas I, II and III are modulators of the signalingprocesses which follow binding of TNF-α to its receptor. In preferredembodiments, the compounds of the invention interrupt the signalingprocess. Without being bound by theory, it is believed that thesecompounds interfere with one or more steps of the signaling cascadewhich includes TNF-α. Thus, these compounds are effective as therapeuticagents for medical conditions in which one or more signaling processesinvolving TNF-α play a role and include, for example, conditions inwhich excessive TNF-α is produced. When selecting substituents for thevarious compounds of Formulas I, II and III, the ordinarily skilledartisan can utilize available information to ensure selection of astable compound with substituents that will enhance the desiredtherapeutic activity. The present compounds can be administered to apatient having a TNF-α mediated medical condition, for example, toinhibit the development of the condition, to inhibit its furtherprogression, and/or to ameliorate the symptoms associated with thecondition.

Thus, in one embodiment, the present invention relates to a method oftreating a TNF-α mediated condition in a patient. The method comprisesthe step of administering to the patient a therapeutically effectiveamount of at least one compound of Formula I, Formula II or Formula III,as described above. The patient can be a human or any animal which issuffering from a TNF-α mediated condition or which is believed to besusceptible to development of a TNF-α mediated condition. Preferably,the patient is a domestic animal, such as a fowl, for example, achicken, or a mammal, for example, a bovine, porcine, canine, feline orequine animal. More preferably, the patient is a human.

The language a “TNF-α mediated condition” is intended to include amedical condition, such as a chronic or acute disease or pathology, orother undesirable physical state, in which a signaling cascade includingTNF-α plays a role, whether, for example, in development, progression ormaintenance of the condition. Examples of TNF-α mediated conditionsinclude, but are not limited to:

-   -   (A) acute and chronic immune and autoimmune pathologies, such as        systemic lupus erythematosus (SLE), rheumatoid arthritis,        thyroidosis, graft versus host disease, scleroderma, diabetes        mellitus, Graves' disease, and the like;    -   (B) infections, including sepsis syndrome, circulatory collapse        and shock resulting from acute or chronic bacterial infection,        acute and chronic parasitic infection, and/or infectious        diseases, whether bacterial, viral or fungal in origin, such as        a HIV or AIDS, and including symptoms of cachexia, autoimmune        disorders, Acquired Immune Deficiency Syndrome, dementia complex        and infections;    -   (C) inflammatory diseases, such as chronic inflammatory        pathologies, including sarcoidosis, chronic inflammatory bowel        disease, ulcerative colitis and Crohn's pathology, and vascular        inflammatory pathologies, such as, disseminated intravascular        coagulation, atherosclerosis and Kawasaki's pathology;    -   (D) neurodegenerative diseases, including, demyelinating        diseases, such as multiple sclerosis and acute transverse        myelitis; extrapyramidal and cerebellar disorders such as        lesions of the corticospinal system; disorders of the basal        ganglia or cerebellar disorders; hyperkinetic movement disorders        such as Huntington's Chorea and senile chorea; drug-induced        movement disorders, such as those induced by drugs which block        CNS dopamine receptors; hypokinetic movement disorders, such as        Parkinson's disease; progressive supranucleo palsy; Cerebellar        and Spinocerebellar Disorders, such as astructural lesions of        the cerebellum; spinocerebellar degenerations, such as spinal        ataxia, Friedreich's ataxia, cerebellar cortical degenerations;        multiple systems degenerations, such as Mencel, Dejerine-Thomas,        Shi-Drager, and Machado-Joseph; systemic disorders, such as        Refsum's disease, abetalipoprotemia, ataxia, telangiectasia, and        mitochondrial multisystem disorder; demyelinating core        disorders, such as multiple sclerosis, acute transverse        myelitis; disorders of the motor unit, such as neurogenic        muscular atrophies, such as anterior horn cell degeneration,        amyotrophic lateral sclerosis, infantile spinal muscular atrophy        and juvenile spinal muscular atrophy; Alzheimer's disease;        Down's Syndrome in middle age; Diffuse Lewy body disease; Senile        Dementia of Lewy body type; Wernicke-Korsakoff syndrome; chronic        alcoholism; Creutzfeldt-Jakob disease; Subacute sclerosing        panencephalitis, Hallerrorden-Spatz disease; and Dementia        pugilistica, or any subset of conditions, symptoms or syndromes        thereof;    -   (E) malignant pathologies involving TNF-α secreting tumors or        other malignancies involving TNF, such as leukemias including        acute, chronic myelocytic, chronic lymphocytic and/or        myelodyspastic syndrome; lymphomas including Hodgkin's and        non-Hodgkin's lymphomas; and malignant lymphomas, such as        Burkitt's lymphoma or Mycosis fungoides; and    -   (F) alcohol-induced hepatitis.

See, e.g., Berkow, et al., eds., The Merck Manual, 16^(th) edition,chapter 11, pp 1380-1529, Merck and Co., Rahway, N.J., (1992), whichreference, and references cited therein, are entirely incorporatedherein by reference.

The language a “therapeutically effective amount” is intended to includean amount which is sufficient to inhibit, totally or partially, theTNF-α mediated condition, prevent its further progression or amelioratethe symptoms associated with the condition. Such an amount, whenadministered prophylactically to a patient thought to be susceptible todevelopment of a TNF-α mediated condition, can also be effective toprevent or lessen the severity of the TNF-α mediated condition.

The compounds of this invention can be administered to the patient bythemselves or in pharmaceutical compositions where they are mixed with asuitable carrier or excipient at doses to treat or ameliorate thesymptoms associated with the TNF-α mediated condition. Mixtures of thesecompounds can also be administered to the patient as a simple mixture orin suitable formulated pharmaceutical compositions. Techniques forformulation and administration of the compounds of the instantapplication can be found in Remington: the Science and Practice ofPharmacy, 19^(th) edition, Mack Publishing Co., Easton, Pa. (1995).

Suitable routes of administration can, for example, include oral,eyedrop, rectal, transmucosal, topical, or intestinal administration;parenteral delivery, including intramuscular, subcutaneous,intramedullary injections, as well as intrathecal, directintraventricular, intravenous, intraperitoneal, intranasal, orintraocular injections.

The compounds of the invention can also be administered in a targeteddrug delivery system, such as, for example, in a liposome coated withendothelial cell-specific antibody.

The invention also relates to the use of a compound of Formula I,Formula II or Formula III, as described above, for the manufacture of amedicament for treating a TNF-α mediated condition.

The pharmaceutical compositions of the present invention can bemanufactured in a manner that is itself known, e.g., by means ofconventional mixing, dissolving, granulating, dragee-making, levigating,emulsifying, encapsulating, entrapping or lyophilizing processes.

Pharmaceutical compositions for use in accordance with the presentinvention thus can be formulated in a conventional manner using one ormore physiologically acceptable carriers comprising excipients andauxiliaries which facilitate processing of the active compounds intopreparations which can be used pharmaceutically. Proper formulation isdependent upon the route of administration chosen.

For injection, the agents of the invention may be formulated in aqueoussolutions, preferably in physiologically compatible buffers such asHanks's solution, Ringer's solution, or physiological saline buffer. Fortransmucosal administration, penetrants appropriate to the barrier to bepermeated are used in the formulation. Such penetrants are generallyknown in the art.

For oral administration, the compounds can be formulated readily bycombining the active compounds with pharmaceutically acceptable carrierswell known in the art. Such carriers enable the compounds of theinvention to be formulated as tablets, pills, dragees, capsules,liquids, gels, syrups, slurries, suspensions and the like, for oralingestion by a patient to be treated. Pharmaceutical preparations fororal use can be obtained by combining the active compound with a solidexcipient, optionally grinding a resulting mixture, and processing themixture of granules, after adding suitable auxiliaries, if desired, toobtain tablets or dragee cores. Suitable excipients are, in particular,fillers such as sugars, including lactose, sucrose, mannitol, orsorbitol, cellulose preparations such as, for example, maize starch,wheat starch, rice starch, potato starch, gelatin, gum tragacanth,methyl cellulose, hydroxypropylmethyl-cellulose, sodiumcarboxymethylcellulose, and/or polyvinylpyrrolidone (PVP). If desired,disintegrating agents may be added, such as the cross-linked polyvinylpyrrolidone, agar, or alginic acid or a salt thereof such as sodiumalginate.

Dragee cores are provided with suitable coatings. For this purpose,concentrated sugar solutions may be used, which may optionally containgum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethyleneglycol, and/or titanium dioxide, lacquer solutions, and suitable organicsolvents or solvent mixtures. Dyestuffs or pigments may be added to thetablets or dragee coatings for identification or to characterizedifferent combinations of active compound doses.

Pharmaceutical preparations which can be used orally include push-fitcapsules made of gelatin, as well as soft, sealed capsules made ofgelatin and a plasticizer, such as glycerol or sorbitol. The push-fitcapsules can contain the active ingredients in admixture with fillersuch as lactose, binders such as starches, and/or lubricants such astalc or magnesium stearate and, optionally, stabilizers. In softcapsules, the active compounds may be dissolved or suspended in suitableliquids, such as fatty oils, liquid paraffin, or liquid polyethyleneglycols. In addition, stabilizers may be added. All formulations fororal administration should be in dosages suitable for suchadministration.

For buccal administration, the compositions may take the form of tabletsor lozenges formulated in a conventional manner.

For administration by inhalation, the compounds for use according to thepresent invention are conveniently delivered in the form of a dry powderinhaler, or an aerosol spray presentation from pressurized packs or anebuliser, with the use of a suitable propellant, e.g.,dichlorodifluoromethane, trichlorofluoromethane,dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In thecase of pressurized aerosol the dosage unit may be determined byproviding a valve to deliver a metered amount. Capsules and cartridgesof gelatin for use in an inhaler or insufflator may be formulatedcontaining a powder mix of the compound and a suitable powder base suchas lactose or starch.

The compounds can be formulated for parenteral administration byinjection, including bolus injection or continuous infusion.Formulations for injection may be presented in unit dosage form, such asin ampoules or in multi-dose containers, with an added preservative. Thecompositions may take such forms as suspensions, solutions or emulsionsin oily or aqueous vehicles, and may contain formulatory agents such assuspending, stabilizing and/or dispersing agents.

Pharmaceutical formulations for parenteral administration includeaqueous solutions of the active compounds in water-soluble form.Additionally, suspensions of the active compounds may be prepared asappropriate oily injection suspensions. Suitable lipophilic solvents orvehicles include fatty oils such as sesame oil, or synthetic fatty acidesters, such as ethyl oleate or triglycerides, or liposomes. Aqueousinjection suspensions may contain substances which increase theviscosity of the suspension, such as sodium carboxymethyl cellulose,sorbitol, or dextran. Optionally, the suspension may also containsuitable stabilizers or agents which increase the solubility of thecompounds to allow for the preparation of highly concentrated solutions.

Alternatively, the active ingredient may be in powder form forconstitution with a suitable vehicle, such as sterile pyrogen-freewater, before use.

The compounds may also be formulated in rectal compositions such assuppositories or retention enemas, e.g., containing conventionalsuppository bases such as cocoa butter or other glycerides.

In addition to the formulations described previously, the compounds mayalso be formulated as a depot preparation. Such long acting formulationsmay be administered by implantation, for example, subcutaneously orintramuscularly or by intramuscular injection. Thus, for example, thecompounds may be formulated with suitable polymeric or hydrophobicmaterials, for example, as an emulsion in an acceptable oil, or ionexchange resins, or as sparingly soluble derivatives, for example, as asparingly soluble salt.

The pharmaceutical compositions can also include suitable solid or gelphase carriers or excipients. Examples of such carriers or excipientsinclude calcium carbonate, calcium phosphate, various sugars, starches,cellulose derivatives, gelatin, and polymers such as polyethyleneglycols.

Pharmaceutical compositions suitable for use in the present inventioninclude compositions wherein the active ingredients are contained in aneffective amount to achieve the intended purpose. More specifically, atherapeutically effective amount, as previously defined, denotes anamount effective to prevent development of or to alleviate the existingsymptoms of the subject being treated. Determination of the effectiveamounts is well within the capability of those skilled in the relevantart.

For any compound used in the method of the invention, thetherapeutically effective dose can be estimated initially from in vitroassays and animal models. For example, a dose can be formulated incellular and animal models to achieve a circulating concentration rangethat includes the IC₅₀ as determined in cellular assays, i.e., theconcentration of the test compound which achieves a half-maximalinhibition of TNF-α activity. In some cases it is appropriate todetermine the IC₅₀ in the presence of 3 to 5% serum albumin, since sucha determination approximates the binding effects of plasma protein onthe compound. Such information can be used to more accurately determineuseful doses in humans. Further, the most preferred compounds forsystemic administration effectively inhibit TNF-α signaling in intactcells at levels that are safely achievable in plasma.

Toxicity and therapeutic efficacy of such compounds can be determined bystandard pharmaceutical procedures in cell cultures or experimentalanimals, e.g., for determining the maximum tolerated dose (MTD) and theED₅₀ (effective dose for 50% maximal response). The dose ratio betweentoxic and therapeutic effects is the therapeutic index and it can beexpressed as the ratio between MTD and ED₅₀. Compounds which exhibithigh therapeutic indices are preferred. The data obtained from thesecell culture assays and animal studies can be used in formulating arange of dosage for use in humans. The dosage of such compounds liespreferably within a range of circulating concentrations that include theED₅₀ with little or no toxicity. The dosage may vary within this rangedepending upon the dosage form employed and the route of administrationutilized. The exact formulation, route of administration and dosage canbe chosen by the individual physician in view of the patient'scondition. (See, e.g., Fingl, et al., 1975, in The Pharmacological Basisof Therapeutics, Chapter 1, p. 1). In the treatment of crises, theadministration of an acute bolus or an infusion approaching the MTD maybe required to obtain a rapid response.

Dosage amount and interval may be adjusted individually to provideplasma levels of the active moiety which are sufficient to maintain thedesired effects, or minimal effective concentration (MEC). The MEC willvary for each compound but can be estimated from in vitro data, e.g.,the concentration necessary to achieve 50-90% inhibition of proteinkinase using the assays described herein. Dosages necessary to achievethe MEC will depend on individual characteristics and route ofadministration. However, HPLC assays or bioassays can be used todetermine plasma concentrations.

Dosage intervals can also be determined using the MEC value. Compoundsshould be administered using a regimen which maintains plasma levelsabove the MEC for 10-90% of the time, preferably between 30-90% and mostpreferably between 50-90% until the desired amelioration of symptoms isachieved. In cases of local administration or selective uptake, theeffective local concentration of the drug may not be related to plasmaconcentration.

The amount of composition administered will be dependent on the subjectbeing treated, on the subject's weight, the severity of the affliction,the manner of administration and the judgment of the prescribingphysician.

Synthetic Strategies

In general, the substituted pyrazole ring of Formula I can be preparedby reaction of a 1,3-dicarbonylalkane with a hydrazine (Scheme I).

When the method in Scheme I is used to construct the compoundsrepresented by Formula I, R₂₈ is ring A and R₂₇ is —X(CH₂)_(n)—Y. R₂₅and R₂₆ are each, independently, —H or a substituted or unsubstitutedalkyl or a substituted or unsubstituted alkoxy. The above reaction wasused to prepare the compounds in Examples 6-10 and 12-15.

The reaction to form the pyrazole ring is carried out in a polarsolvent, such as water, an alcohol or an ether. Preferably, the reactionis carried out in an alcohol, such as ethanol. The reaction temperatureis about 35° C. to about 150° C., preferably about 70° C. to about 90°C. Typically, the reaction is carried out at the reflux temperature ofthe solvent used.

In compounds which can be represented by Formula I, ring A can be apyridine, a pyrimidine or a triazine ring. Substituted2-hydrazinopyrimidines can be prepared by addition of thiourea to3-(N,N-dimethyamino)prop-2-en-1-one in the presence of a base, followedby in situ methylation of the cyclic thiourea formed (see Scheme II,step 1). The resultant 4-substituted 2-methylsulfanylpyrimidine istreated with hydrazine and heat to form a 4-substituted2-hydrazinopyrimidines (see Scheme II, step 2).

The reaction depicted in Scheme II was used in Example 5 to prepare theintermediate, 2-hydrazino-4-(pyridin-2-yl)-pyrimidine. This intermediatewas reacted with a 1,3-dicarbonylalkane via the method shown in Scheme Ito prepare a substituted pyrazole in Examples 6-9.

The reaction to form substituted 2-methylsulfanylpyrimidine (see SchemeII, step 1) is carried out in a polar solvent, such as water, an alcoholor an ether. Preferably, the reaction is carried out in an alcohol, suchas ethanol, using an alkaline or alkaline earth metal alkoxide as thebase. The reaction temperature is about 35° C. to about 150° C.,preferably about 70° C. to about 90° C. Typically, the reaction iscarried out at the reflux temperature of the solvent used. The reactionis usually allowed to proceed for about 1 hour to about 24 hours,preferably about 2 to about 5 hours at an elevated temperature, then thereaction is cooled and a primary haloalkane, preferably an iodoalkane,is added to alkylate the thiol group.

The substituted 2-methylsulfanylpyrimidine is then treated withhydrazine at elevated temperatures to form substituted2-hydrazinopyrimidine. The reaction can be carried out in a polarsolvent, such as water, an alcohol or an ether. Typically, the reactionis carried out in water having about 35% (vol/vol) hydrazine. Thereaction temperature is about 80° C. to about 110° C., preferably, about100° C.

Substituted 2-hydrazinotriazines can be prepared by reacting2-amidopyridine or 2-amidoquinoxaline with analkoxy-bis-(dimethylamino)methane, such astert-butoxy-bis-(dimethylamino)methane, at a temperature of about 80° C.to about 170° C., preferably at about 145° C. to about 160° C. for about1 hour to about 5 hours, preferably about 2 hours to about 4 hours toyield a first intermediate. The reaction is typically carried out in apolar aprotic solvent such as dimethylformamide. The 2-amidopyridine or2-amidoquinoxaline is typically present in a concentration of about 0.2M to about 1 M and the alkoxy-bis-(dimethylamino)methane is present inabout 1.5 equivalents to about 3 equivalents in relation to the2-amidopyridine or 2-amidoquinoxaline (Scheme III, step 1a).

The first intermediate is then contacted with thiourea and a base in apolar solvent such as water, an alcohol or an ether. Preferably, thereaction is carried out in an alcohol, such as ethanol, using analkaline or alkaline earth metal alkoxide as the base. The reactiontemperature is about 35° C. to about 150° C., preferably about 70° C. toabout 90° C. The reaction is usually allowed to proceed for about 1 hourto about 24 hours, preferably about 2 to about 5 hours at an elevatedtemperature to form a 4-substituted-2-thiotriazine (see Scheme III, step1b). The 4-substituted-2-thiotriazine is then alkylated and substitutedwith hydrazine (see Scheme III, steps 2 and 3) to form a substituted2-hydrazinotriazine via the method described above for formingsubstituted 2-hydrazinopyrimidine.

2-Hydrazinotriazine can react with a 1,3-dicarbonylalkane to form apyrazole ring as shown in the method of Scheme I. Examples 12-14 wereprepared via the method of Schemes III followed by the method of SchemeI.

When ring A is a pyridine ring, the pyrazole ring can be formed first byreaction of hydrazine with 1,3-dicarbonylalkane via the method shown inScheme I where R₂₈ is a hydrogen. The pyrazole ring can then be added tothe pyridine ring via a substitution reaction (see Scheme IV).

In Scheme IV, R₂₉ is a leaving group, such as a halogen. The pyrazole iscontacted with a base that is sufficiently strong to deprotonate thepyrazole for about 3 minutes to about 30 minutes prior to addition ofthe substituted pyridine. Typically, a hydride salt, such as sodiumhydride is used. After addition of the substituted pyridine the reactionis heated for about 6 hours to about 24 hours at a temperature of about80° C. to about 120° C. to form a 6-substituted2-(pyrazol-1-yl)pyridine. The reaction is typically carried out in anaprotic solvent such as an ether. Example 10 was prepared using themethod of Scheme I followed by the method of Scheme IV.

The compounds listed in Tables 2 and 4 were prepared using the methodsof Schemes I-IV.

The substituted triazole ring of Formula II can be prepared by reactionof a N-thiocarbonylamide with a hydrazine (see Scheme V).

When the reaction in Scheme V is used to form the compounds of FormulaII, R₂₄ is a 4-(pyridin-2-yl)triazin-2-yl. The reaction is carried outin an acidic buffer, such as carboxylic acid and the salt of thecarboxylic acid, for example acetic acid and sodium acetate. An organicsolvent can also be present, such as an ether. The reaction is heated toabout 70° C. to about 110° C. for about 6 hours to about 24 hours.Example 11 is representative of the method depicted in Scheme V.

The compounds in Table 3 were prepared using the methods of Schemes IIIand V.

The compounds represented by Formula III can be formed via an Ugireaction, as shown in Scheme VI.

For reviews of Ugi reactions see Gross and Meienhofer, The Peptides,vol. 2, pp. 365-381, Academic Press, New York, (1980); Intra-Sci. Chem.Rep. (1971), 5:229-261; Rec. Chem. Prog. (1969), 30:289-311; and Eberle,et al., Tetrahedron (1978), 34:977, the entire teachings of which areincorporated herein by reference. The starting materials for the Ugireaction, i.e., an isonitrile, a carboxylic acid, an aldehyde or aketone and an amine, are mixed together in a polar solvent such as analcohol, preferably methanol or ethanol. The reaction is heated to about40° C. to about 80° C. for about 6 hours to about 24 hours. Examples 1and 2 are representative of compounds prepared using an Ugi reaction.

A limitation of the Ugi reaction is that the terminal nitrogen atom,i.e., the nitrogen substituted with R₈) is monosubstituted. Therefore, asecond method of forming the compounds represented by Formula III wasdeveloped wherein the terminal nitrogen can be mono- or disubstituted(see Scheme VII).

In Scheme VII, R₂₉ is a leaving group, such as a halide, and R₃₀ is asubstituted or unsubstituted aryl or a substituted or unsubstitutedalkyl. The starting materials for the first step are mixed together inabout equal molar amounts in a nonpolar, aprotic solvent such asmethylene chloride, or an ether, for about 0.5 hours to about 6 hours,preferably about 1 hour to form an acetoxyamide. This is followed byhydrolysis with lithium hydroxide to form an α-hydroxyamide.

In step 2, the α-hydroxyamide formed in step 1 is dissolved in anaprotic solvent, such as an ether, at a concentration of about 0.2 M toabout 0.4 M and is treated with about 1.1 equivalents to about 1.5equivalents, preferably about 1.2 equivalents, of a strong base, such asa hydride salt, for example, sodium hydride, for about 2 minutes toabout 20 minutes, followed by addition of about 1.1 equivalents to about1.5 equivalents, preferably about 1.2 equivalents, of an arylsulfonylhalide or alkylsulfonyl halide. After addition of the arylsulfonylhalide or alkylsulfonyl halide, the reaction is allowed to proceed forabout 1 hour to about 6 hours, then about 3 equivalents to about 5equivalents, preferably about 4 equivalents, of a primary amine is addedto the reaction mixture accompanied by addition of a polar solvent suchas an alcohol. After addition of the primary amine, the reaction isallowed to proceed for about 6 hours to about 24 hours.

The product of step 2 is treated with about 1.1 equivalents to about 1.8equivalents of an acid halide in a non-polar solvent, such as ahalogenated alkane or an ether in the presence of a base, such as atertiary amine, for about 1 hour to about 4 hours at a temperature ofabout 30° C. to about 80° C. to form the desired product. Example 4 isrepresentative of this method.

A third method of forming the compounds represented by Formula III wasdeveloped wherein the stereochemistry of the carbon substituted with R₉can be controlled (see Scheme VIII).

In Scheme VIII, R₂₉ is a leaving group, such as a halogen and R₃₁ is anamine protecting group. Methods for protecting amines can be found inGreene, et al., Protecting Groups in Organic Synthesis, 2^(nd) Edition,John Wiley & Sons, Inc., (1991), pp. 309-405, the entire teachings ofwhich are incorporated herein by reference. The starting material forthe synthesis depicted in Scheme VIII can be a natural or unnaturalamino acid wherein the amine group is protected. Preferred protectinggroups for the amine are 9-fluorenylmethyl carbamate and t-butylcarbamate. When a chiral amino acid is the starting material, the chiralcenter at the α-carbon is retained through out the synthesis.

In step 1 of Scheme VIII, the carboxylic acid of the starting materialis transformed into an activated ester by methods known to those skilledin the art. The activated ester is then contacted with a primary amineto form an amide. In step 2, a Mitsunobu reaction is used to displace ahydroxyl group of R₁₀—OH with the protected amine of the product formedin step 1 by treating the alcohol with triphenylphosphine anddi-tert-butyl azodicarboxylate in the presence of the product of step 1.Fukuyama T., et al., Tetrahedron Letters (1995), 36:6373. The amine isthen deprotected in step 3 and reacted with an acid halide as describedabove for step 3 of synthetic Scheme VII. Example 3 is representative ofthis method.

The compounds of Table I were formed using the methods of SchemesVI-VIII.

EXAMPLES I. Synthetic Methods Example 12-Benzoyl-N-[(2,2-diphenyl-ethylcarbamoyl)-phenyl-methyl]-N-[2-(1H-imidazol-4-yl)-ethyl]-benzamide(Compound 5) Step 1) Preparation of 2,2-Diphenylethylisocyanide

A stirred suspension of 2,2-diphenylethylamine (7.00 g, 35.5 mmol) inethyl formate (25 mL) was heated at reflux overnight. The reaction wasconcentrated to afford the corresponding formamide as an off-whitesolid. The crude product was taken up in methylene chloride (50 mL) anddiisopropylamine (13.3 mL, 94.9 mmol) and cooled in an ice bath. Whilestirring, phosphorus oxychloride (5.0 mL, 54 mmol) was added. One hourlater, the ice bath was removed and aqueous sodium carbonate solution(75 mL), water (50 mL) and methylene chloride (50 mL) were added. Themixture was stirred vigorously for 1 hour. The organic layer was washedwith water, dried (sodium sulfate) and concentrated. The resulting crudeproduct was filtered through a plug of silica (methylene chloride) toafford 6.62 g (90%) of product as an off-white solid: ¹H NMR (CDCl₃) δ7.38-7.18 (m, 10H), 4.35 (t, J=7.5 Hz, 1H), 3.98 (d, J=7.3, 2H) ppm.

Step 2) Preparation of Compound 5

A solution of histamine (1.11 g, 10.0 mmol), 2-benzoylbenzoic acid (2.26g, 10.0 mmol), benzaldehyde (1.06 g, 10.0 mmol) and the product of step1 (2.07 g, 10.0 mmol) in methanol was heated at reflux overnight. Thereaction was concentrated and the residue partitioned between ethylacetate and aqueous sodium bicarbonate solution. The organic layer wasdried (sodium sulfate) and concentrated to afford a foamy light brownsolid. Flash chromatography over silica (methylene chloride/methanol)afforded 2.99 g (47%) of the product as a beige solid: ¹H NMR (CDCl₃) δ7.88-6.84 (m, 25H), 6.65-6.25 (m, 2H), 5.66-5.30 (m, 1H), 4.44-4.20 (m,1H), 4.17-3.84 (m, 2H), 3.47-3.03 (m, 2H), 2.69-2.18 (m, 2H) ppm. MS(ESI) m/z 634 (M+H⁺).

Example 2N-[(4-Cyano-phenyl)-(2,2-diphenyl-ethylcarbamoyl)-methyl]-2-(2,2-dimethyl-propionyl)-N-[2-(1H-imidazol-4-yl)-ethyl]-benzamide(Compound 54)

A solution of histamine (0.100 g, 0.900 mmol), 2-pivaloylbenzoic acid(0.186 g, 0.902 mmol), 3-cyanobenzaldehyde (0.118 g, 0.900 mmol) and theproduct of step 1 of example 1 (0.187 g, 0.902 mmol) in methanol washeated at reflux overnight. The reaction was concentrated and theresidue partitioned between ethyl acetate and aqueous sodium bicarbonatesolution. The organic layer was dried (sodium sulfate) and concentratedto afford 0.521 g (91%) of crude product as an foamy amber solid. Thismaterial was used for biological testing without further purification.

Example 32-Benzoyl-N-[(1′R)-1′(2,2′-diphenyl-ethylcarbamoyl)-2′-phenyl-ethyl]-N-[2-(1H-imidazol-4-yl)-ethyl]-benzamide(Compound 47) Step 1) Preparation of(2R)-2-Amino-N-(2,2-diphenyl-ethyl)-3-phenyl-propionamide

To a stirred solution of N-(tert-butoxycarbonyl)-D-phenylalanine (11.0g, 41.5 mmol) in chloroform (125 mL) was added1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (7.95 g,41.5 mmol) and 1-hydroxybenzotriazole (5.60 g, 41.4 mmol). After 5minutes, 2,2-diphenylethylamine (7.44 g, 37.7 mmol) was added. Thereaction was stirred for 2 hours, diluted with chloroform (100 mL) andwashed with aqueous sodium bicarbonate solution. The organic layer wasdried (sodium sulfate) and concentrated to afford an off-white solid.Trituration with ether afforded the crude amide. This material was takenup in methylene chloride (100 mL) and treated with trifluoroacetic acid(45 mL, 580 mmol). After stirring for 15 minutes, the reaction wasconcentrated and the residue was partitioned between ethyl acetate andaqueous sodium bicarbonate. The organic layer was dried (sodium sulfate)and concentrated to afford 12.4 g (111% yield inflated by entrainedresidual solvent) of product as a colorless gum: ¹H NMR (CDCl₃) δ7.34-6.95 (m, 15H), 4.10 (t, J=8.2, 1H), 3.88-3.79 (m, 2H), 3.68-3.61(m, 1H), 3.49 (br s, 2H), 3.13-3.03 (m, 1H), 2.70-2.59 (m, 1H) ppm.

Step 2) Preparation of(2R)—N-(2,2-Diphenyl-ethyl)-2-(2-nitro-benzenesulfonylamino)-3-phenyl-propionamide

To a stirred solution of the product of step 1 (12.3 g, 35.6 mmol) inmethylene chloride (200 mL) was added 2-nitrobenzenesulfonyl chloride(9.00 g, 40.6 mmol) followed by triethylamine (6.4 mL, 46 mmol). After30 minutes the reaction was concentrated and the residue partitionedbetween ethyl acetate and aqueous sodium bicarbonate solution. Theorganic layer was dried (sodium sulfate) and concentrated. The resultingamber foam was purified by flash chromatography over silica(hexane/ethyl acetate) to afford 14.0 g (74%) of product as a colorlessfoam: ¹H NMR (CDCl₃) δ 7.95-7.87 (m, 1H), 7.76-7.62 (m, 3H), 7.35-7.15(m, 10H), 7.06-6.94 (m, 3H), 6.90-6.83 (m, 2H), 6.42 (t, J=5.6, 1H),5.80 (d, J=5.8, 1H), 4.12 (t, J=8.1, 1H), 4.01-3.82 (m, 3H), 3.18-3.08(m, 1H), 2.68-3.57 (m, 1H) ppm.

Step 3) Preparation of(2R)—N-(2,2-Diphenyl-ethyl)-2-{(2-nitro-benzenesulfonyl)-[2-(1-trityl-1H-imidazol-4-yl)-ethyl]-amino}-3-phenyl-propionamide

To a stirred solution of the product of step 2 (3.40 g, 6.42 mmol) and2-(1-trityl-1H-imidazol-4-yl)-ethanol (2.73 g, 7.70 mmol) in methylenechloride (40 mL) was added triphenylphosphine (2.19 g, 8.35 mmol)followed by di-tert-butyl azodicarboxylate (1.77 g, 7.69 mmol). After 3hours the reaction was concentrated and the residue partitioned betweenethyl acetate and aqueous sodium bicarbonate solution. The organic layerwas dried (sodium sulfate) and concentrated to yield an amber gum. Flashchromatography over silica (methylene chloride/methanol) afforded 5.42 g(97%) of partially purified product (contaminated with di-tert-butylhydrazodiformate) as a pale yellow foam: ¹H NMR (CDCl₃) δ 7.81-7.74 (m,1H), 7.64-7.55 (m, 1H), 7.53-7.44 (m, 2H), 7.41-7.03 (m, 31H), 6.69 (t,J=5.6, 1H), 6.53 (s, 1H), 4.48 (t, J=8.1, 1H), 4.08 (t, J=8.1, 1H),3.84-3.58 (m, 4H), 3.44-3.34 (m, 1H), 2.91-3.80 (m, 1H), 2.78-2.68 (m,1H), 2.63-2.51 (m, 1H) ppm.

Step 4) Preparation of(2R)—N-(2,2-Diphenyl-ethyl)-3-phenyl-2-[2-(1-trityl-1H-imidazol-4-yl)-ethylamino]-propionamide

To a stirred solution of the product of step 3 (5.42 g, 6.26 mmol) inN,N-dimethylformamide (17 mL) was added mercaptoacetic acid (1.1 mL,15.8 mmol) followed by lithium hydroxide hydrate (1.31 g, 31.2 mmol).The mixture was stirred for 3 hours and then partitioned between ethylacetate and aqueous sodium bicarbonate solution. The organic layer wascombined with a second ethyl acetate extract, dried (sodium sulfate) andconcentrated to furnish an amber oil. Flash chromatography over silica(methylene chloride/methanolic ammonia solution) provided 2.50 g (59%)of product as a colorless tacky foam: ¹H NMR (CDCl₃) δ 7.54 (t, J=5.5,1H), 7.38-7.01 (m, 31H), 6.33 (s, 1H), 4.16 (t, J=8.3, 1H), 4.02-3.92(m, 1H), 3.83-3.74 (m, 1H), 3.22-3.15 (m, 1H), 3.12-3.04 (m, 1H),2.51-2.18 (m, 5H) ppm.

Step 5) Preparation of2-Benzoyl-N-[(1′R)-1-(2′,2′-diphenyl-ethylcarbamoyl)-2′-phenyl-ethyl]-N-[2-(1-trityl-1H-imidazol-4-yl)-ethyl]-benzamide

2-Benzoylbenzoic acid (1.33 g, 5.88 mmol) was taken up in a 2M solutionof oxalyl chloride in methylene chloride (4.5 mL, 9.0 mmol). One drop ofN,N-dimethylformamide was added and the frothy mixture was stirred for 2hours before concentrating. To the residue was added a solution of theproduct of step 4 (2.50 g, 3.67 mmol) in chloroform (30 mL) andtriethylamine (1.05 mL, 7.53 mmol). The mixture was heated at reflux for2.5 hours. At this time, more triethylamine (0.50 mL, 3.6 mmol) wasadded and the reaction was heated for additional 30 minutes. Thereaction was then concentrated and the residue was partitioned betweenethyl acetate and aqueous sodium bicarbonate solution. The organic layerwas dried (sodium sulfate) and concentrated to afford an amber foam.This material was filtered through silica (methylene chloride/methanolicammonia solution) to afford 1.44 g (44%) of a pale amber foam which wasused without further purification in the next step.

Step 6) Preparation of Compound 47

A stirred solution of the product of step 5 (1.44 g, 1.62 mmol) in 4:1acetic acid/water (7 mL) was heated in a water bath (90° C.) for 30minutes. The reaction solution was then cooled and partitioned betweenethyl acetate and aqueous sodium carbonate solution. The organic layerwas combined with a second ethyl acetate extract, dried (sodium sulfate)and concentrated to afford an amber foam. Flash chromatography oversilica (methylene chloride/methanol) afforded 0.62 g (59%) of product asa foamy pale amber solid. Comparisons by thin layer chromatography and¹H NMR spectroscopy indicated that this material is identical to thecorresponding racemic Ugi product, compound 43. Analytical chiral HPLC(ChiralPack AD, 4.6×250 mm; eluant: 0.1% diethylamine in 88:12hexane/ethanol; flow: 1 mL/min; detection: 260 nM) indicated that thematerial is a single enantiomer within detection limits: ¹H NMR (CDCl₃)δ 8.35 (br s, 1H), 7.90-6.94 (m, 27H), 6.78-6.57 (m, 1H), 6.39-6.17 (m,1H), 4.65-3.72 (m, 4H), 3.64-2.70 (m, 5H), 2.46-2.18 (m, 1H) ppm. MS(ESI) m/z 648 (M+H⁺).

Example 42-Benzoyl-N-{[(2,2-diphenyl-ethyl)-methyl-carbamoyl]-phenyl-methyl}-N-[2-(1H-imidazol-4-yl)-ethyl]-benzamide(Compound 26) Step 1) PreparationN-(2,2-Diphenyl-ethyl)-2-hydroxy-N-methyl-2-phenyl-acetamide

To a stirred solution of (2,2-diphenylethyl)methylamine (4.97 g, 23.5mmol) in chloroform (50 mL) was added 0-acetylmandelic chloride (5.3 mL,24 mmol) followed by triethylamine (4.0 mL, 29 mmol). The reaction wasstirred for 1 hour and then concentrated. The residue was partitionedbetween ethyl acetate and aqueous sodium bicarbonate solution. Theorganic layer was dried (sodium sulfate) and concentrated to afford acolorless gum. The crude amide was taken up in 1:1tetrahydrofuran/methanol (80 mL) and treated with 1 N aqueous lithiumhydroxide (30 mL, 30 mmol). The mixture was stirred for 2 hours andconcentrated to remove the organic solvents. The remaining aqueoussolution was diluted with water and extracted twice with ethyl acetateand once with methylene chloride. The combined organic extracts weredried (sodium sulfate) and concentrated to afford an off-white solid.This material was triturated with diethyl ether to afford 7.59 g (93%)of product as a white solid: ¹H NMR (CDCl₃) δ 7.43-6.98 (m, 15H), 5.07(s, 1H), 4.57-4.44 (m, 1H), 4.36-4.28 (m, 1H), 3.76-3.64 (m, 1H), 2.46(s, 3H) ppm.

Step 2) Preparation ofN-(2,2-Diphenyl-ethyl)-2-[2-(1H-imidazol-4-yl)-ethylamino]-N-methyl-2-phenyl-acetamide

To stirred solution of the product of step 1 (1.50 g, 4.34 mmol) intetrahydrofuran (15 mL) was added a 60% dispersion of sodium hydride inmineral oil (0.210 g, 5.25 mmol). After gas evolution ceased,p-toluenesulfonyl chloride (0.990 g, 5.20 mmol) was added and themixture was stirred for 2 hours. At this time, histamine (1.93 g, 17.4mmol) was added followed by methanol (10 mL). The reaction was stirredovernight and concentrated. The residue was partitioned between ethylacetate and aqueous sodium carbonate solution. The organic layer wasdried (sodium sulfate) and concentrated to afford a pasty white solid.Flash chromatography over silica (methylene chloride/methanolic ammoniasolution) provided 1.08 g (57%) of product as a colorless tacky foam: ¹HNMR (CDCl₃) δ 7.52-7.43 (m, 1H), 7.40-7.00 (m, 15H), 6.78-6.69 (m, 1H),4.36-3.97 (m, 4H), 2.92-2.20 (m, 7H) ppm.

Step 3) Preparation of Compound 26

2-Benzoylbenzoic acid (0.341 g, 1.51 mmol) was taken up in a 2M solutionof oxalyl chloride in methylene chloride (1.5 mL, 3.0 mmol). One drop ofN,N-dimethylformamide was added and the frothy mixture was stirredovernight and concentrated. A solution of the crude 2-benzoylbenzoylchloride in methylene chloride (4 mL) followed by triethylamine (0.25mL, 1.8 mmol) was added to the product of step 2 (0.220 g, 0.502 mmol).The mixture was heated at reflux for 2 hours and concentrated. Theresidue was taken up in 5:1 methanol/1 N aqueous lithium hydroxide (12mL) and stirred for 1 hour. The reaction was again concentrated and theresidue was partitioned between ethyl acetate and aqueous sodiumbicarbonate solution. The organic layer was dried (sodium sulfate) andconcentrated to afford a foamy amber solid. Flash chromatography oversilica (methylene chloride/methanol) provided 0.229 g (71%) of productas a foamy pale amber solid: ¹H NMR (CDCl₃) δ 7.91-6.92 (m, 26H),6.54-6.39 (m, 1H), 6.34-6.06 (m, 1H), 4.83-4.31 (m, 2H), 3.97-2.82 (m,3H), 2.70-2.39 (m, 3H), 2.35-2.09 (m, 2H) ppm. MS (ESI) m/z 648 (M+H⁺).

Example 5 2-Hydrazino-4-(Pyridin-2-yl)-Pyrimidine Step 1) Preparation of2-Methylsulfanyl-4-pyridin-2-yl-pyrimidine Synthetic Intermediate Usedin Examples 6, 7, 8 & 15

To a stirred solution of sodium ethoxide (freshly prepared from sodiummetal (5.11 g, 222 mmol) and ethanol (350 mL)) was added3-dimethylamino-1-pyridin-2-yl-propenone (28.0 g, 159 mmol) and thiourea(12.9 g, 170 mmol). The solution was heated to reflux and stirred for 3hours. The solution was allowed to cool to room temperature, and theniodomethane (13.8 mL, 222 mmol) was added dropwise over 10 minutes. Thesolution was stirred at room temperature for 2 hours, and then dilutedwith saturated ammonium chloride solution (250 mL) and water (250 mL).The suspension was extracted with ethyl ether (200 mL×3), and thecombined organic extracts washed with saturated sodium thiosulfatesolution (150 mL) and brine (150 mL). The organic phase was dried(magnesium sulfate), filtered, and concentrated to provide 30.1 g (96%)of the product as a brown oil: ¹H NMR (CDCl₃) δ 8.70-8.69 (m, 1H), 8.64(d, J=5.2 Hz, 1H), 8.48 (d, J=7.9 Hz, 1H), 8.01 (d, J=5.2 Hz, 1H),7.87-7.82 (m, 1H), 7.41-7.38 (m, 1H), 2.65 (s, 3H) ppm. ¹³C NMR (CDCl₃)δ 172.6, 163.1, 158.5, 153.9, 149.7, 137.3, 125.7, 122.0, 112.6, 14.5ppm.

Step 2) Preparation of (4-Pyridin-2-yl-pyrimindin-2-yl)-hydrazine

To hydrazine hydrate (90 mL) was added2-methylsulfanyl-4-pyridin-2-yl-pyrimidine (30.0 g, 148 mmol). Thesolution was heated to reflux and stirred for 17 hours. The solution wasallowed to cool to room temperature, and a yellow precipitate formed.The solids were removed by filtration, washed with water, and dried toprovide 22.5 g (81%) of the product as yellow micro needles: ¹H NMR(DMSO-d₆) δ 8.68-8.67 (m, 1H), 8.45-8.41 (m, 2H), 8.29 (s, 1H),7.98-7.93 (m, 1H), 7.52-7.48 (m, 2H), 4.28 (s, 2H) ppm. ¹³C NMR(DMSO-d₆) δ 164.6, 162.6, 159.3, 153.9, 149.4, 125.5, 121.0, 106.0 ppm.

Example 62-[4-(4-Isopropyl-phenoxy)-3,5-dimethyl-pyrazol-1-yl]-4-pyridin-2-yl-pyrimidine(Compound 97) Step 1) Preparation of3-(4-Isopropyl-phenoxy)-pentane-2,4-dione

To a refluxing solution of 4-isopropylphenol (0.548 g, 4.02 mmol) andrhodium(II) acetate (ca. 15 mgs) in benzene (10 mL) was added a solutionof 3-diazo-pentane-2,4-dione (0.507 g, 4.02 mmol) in benzene (20 mL)over 40 minutes. The reaction mixture was allowed to cool to roomtemperature and was concentrated to afford a green oil. Flashchromatography over silica (ethyl acetate/hexanes) provided 0.371 g(39%) of the product as a white solid: ¹H NMR (CDCl₃) δ 14.4 (s, 1H),7.15 (d, J=8.6 Hz, 2H), 6.82 (d, J=8.6 Hz, 2H), 2.87 (sept, J=7.0 Hz,1H), 2.04, (s, 6H), 1.23 (d, J=7.0 Hz, 6H) ppm.

Step 2) Preparation of Compound 97

To a solution of 3-(4-isopropyl-phenoxy)-pentane-2,4-dione (0.371 g,1.58 mmol) in ethanol (20 mL) was added(4-pyridin-2-yl-pyrimindin-2-yl)-hydrazine (0.296 g, 1.58 mmol) andp-toluenesulfonic acid monohydrate (ca. 10 mgs). The solution was heatedto reflux and stirred for 14 hours. The reaction mixture was allowed tocool to room temperature, diluted with water (100 mL), and extractedwith ethyl acetate (100 mL). The organic phase was washed with saturatedsodium hydrogencarbonate solution (50 mL) and brine (50 mL), dried(magnesium sulfate), filtered, and concentrated to afford a yellowsolid. Flash chromatography over silica (methanol/methylene chloride)provided 0.421 g (69%) of the product as a light yellow solid: ¹H NMR(CDCl₃) δ 8.93 (d, J=5.1 Hz, 1H), 8.75-8.74 (m, 1H), 8.46 (d, J=7.9 Hz,1H), 8.24 (d, J=5.1 Hz, 1H), 7.88 (dt, J=7.9, 1.7 Hz, 1H), 7.46-7.42 (m,1H), 7.14 (d, J=8.7 Hz, 2H), 6.88 (d, J=8.7 Hz, 2H), 2.88 (sept, J=6.9Hz, 1H), 2.69 (s, 3H), 2.23 (s, 3H), and 1.23 (d, J=6.9 Hz, 6H) ppm. ¹³CNMR (CDCl₃) δ 164.3, 160.2, 157.5, 156.4, 153.4, 149.7, 145.4, 142.5,137.9, 137.2, 133.1, 127.4, 125.7, 122.0, 114.7, 114.0, 33.3, 24.1,12.7, 11.3 ppm.

Example 72-{4-[4-(4-Benzyl-piperazin-1-ylmethyl)-phenoxy]-3,5-dimethyl-pyrazol-1-yl}-4-pyridin-2-yl-pyrimidine(Compound 104) Step 1) Preparation of{4-[3,5-Dimethyl-1-(4-pyridin-2-yl-pyrimidin-2-yl)-1H-pyrazol-4-yloxy]-phenyl}-methanol

To a solution of 3-(4-acetoxymethyl-phenoxy)-pentane-2,4-dione (1.24 g,4.69 mmol) in ethanol (30 mL) was added(4-pyridin-2-yl-pyrimindin-2-yl)-hydrazine (0.877 g, 4.69 mmol) andp-toluenesulfonic acid monohydrate (about 10 mgs). The reaction mixturewas heated to reflux and stirred for 14 hours. The solution was allowedto cool to room temperature, and water (100 mL) was added. The resultingprecipitate was removed by filtration, washed with water, and dried toprovide a white solid. This crude material was dissolved in 5:1methanol/water (60 mL) and treated with excess potassium carbonate. Thereaction mixture was stirred at room temperature for 2 hours and thendiluted with water (100 mL). The precipitate that formed was isolated byfiltration, washed with water, and dried to provide 1.53 g (87%) of theproduct as a white solid. ¹H NMR analysis was consistent with theassigned structure.

Step 2) Preparation of4-[3,5-Dimethyl-1-(4-pyridin-2-yl-pyrimidin-2-yl)-1H-pyrazol-4-yloxy]-benzaldehyde

To a solution of{4-[3,5-dimethyl-1-(4-pyridin-2-yl-pyrimidin-2-yl)-1H-pyrazol-4-yloxy]-phenyl}-methanol(1.53 g, 4.10 mmol) in chloroform (80 mL) was added the Dess-Martinperiodinane (2.36 g, 5.56 mmol). The reaction mixture was stirred atroom temperature for 3 hours and then concentrated to provide a whitepaste. Flash chromatography over silica (methanol/methylene chloride)provided 1.37 g (90%) the product as a white solid. ¹H NMR analysis wasconsistent with assigned structure.

Step 3) Preparation of Compound 104

To a solution of4-[3,5-dimethyl-1-(4-pyridin-2-yl-pyrimidin-2-yl)-1H-pyrazol-4-yloxy]-benzaldehyde(0.545 g, 1.47 mmol) and 1-benzylpiperazine (0.310 g, 1.76 mmol) indichloroethane (10 mL) was added sodium triacetoxyborohydride (0.436 g,2.06 mmol). The reaction mixture was stirred at room temperature for 17hours. The solution was diluted with methylene chloride (50 mL) andwashed with saturated sodium hydrogencarbonate (2×30 mL) and brine (50mL). The organic phase was dried (magnesium sulfate), filtered, andconcentrated to afford a yellow oil. Flash chromatography over silica (2M ammonia in methanol/methylene chloride) provided 0.183 g (23%) of theproduct as a white foam: ¹H NMR (CDCl₃) δ 8.94-8.93 (m, 1H), 8.75 (d,J=3.9 Hz, 1H), 8.46 (d, J=7.9 Hz, 1H), 8.25 (d, J=3.9 Hz, 1H), 7.90-7.87(m, 1H), 7.46-7.43 (m, 1H), 7.31-7.21 (m, 7H), 6.89 (d, J=8.5 Hz, 2H),3.52 (s, 2H), 3.47 (s, 2H), 2.68 (s, 3H), 2.48 (br s, 8H), 2.21 (s, 3H)ppm.

Example 82-[4-(4-Chloro-benzylsulfanyl)-3,5-dimethyl-pyrazol-1-yl]-4-pyridin-2-yl-pyrimidine(Compound 119) Step 1) Preparation of3-(4-Chloro-benzylsulfanyl)-pentane-2,4-dione

To a solution of 3-chloro-pentan-2,3-dione (1.94 g, 14.4 mmol) inethylene glycol dimethyl ether (50 mL) was added sodiumhydrogencarbonate (ca. 15 g) and 4-chlorobenzyl mercaptan (2.29 g, 14.4mmol). The suspension was heated to reflux and stirred for 3 hours. Thereaction mixture was allowed to cool to room temperature, and water (200mL) was added. The mixture was extracted with ethyl ether (100 mL×2),and the combined organic extracts washed with brine (150 mL). Theorganic phase was dried (magnesium sulfate), filtered, and concentratedto afford a yellow oil. Flash chromatography over silica (ethylacetate/hexanes) provided 2.48 g (67%) of the product as a white solid:¹H NMR (CDCl₃) δ 7.26 (d, J=8.4 Hz, 2H), 7.06 (d, J=8.4 Hz, 2H), 3.60(s, 2H), 2.13 (s, 6H) ppm.

Step 2) Preparation of Compound 119

To a solution of 3-(4-chloro-benzylsulfanyl)-pentane-2,4-dione (1.15 g,4.48 mmol) in ethanol (60 mL) was added(4-pyridin-2-yl-pyrimindin-2-yl)-hydrazine (0.838 g, 4.48 mmol) andp-toluenesulfonic acid monohydrate (ca. 20 mgs). The solution was heatedto reflux and stirred for 5 hours. The reaction mixture was allowed tocool to room temperature, and water (200 mL) was added. The precipitatethat formed was isolated by filtration and recrystallized from aqueousmethanol to provide 1.36 g (74%) of the product as white needles: ¹H NMR(CDCl₃) δ 8.94-8.92 (m, 1H), 8.75-8.73 (m, 1H), 8.41 (d, J=7.9 Hz, 1H),8.28-8.26 (m, 1H), 7.91 (t, J=7.9 Hz, 1H), 7.46-7.43 (m, 1H), 7.20 (d,J=8.0 Hz, 2H), 6.89 (d, J=8.0 Hz, 2H), 3.66 (s, 2H), 2.46 (s, 3H), 2.28(s, 3H) ppm. ¹³C NMR (CDCl₃) δ 164.6, 160.5, 157.4, 155.2, 153.5, 149.8,147.3, 137.6, 137.2, 133.1, 130.6, 128.7, 126.1, 122.2, 114.7, 111.3,40.1, 14.2, 12.6 ppm.

Example 92-[4-(4-Chloro-phenylmethanesulfonyl)-3,5-dimethyl-pyrazol-1-yl]-4-pyridin-2-yl-pyrimidine(Compound 124)

To a suspension of2-[4-(4-chloro-benzylsulfanyl)-3,5-dimethyl-pyrazol-1-yl]-4-pyridin-2-yl-pyrimidine(0.920 g, 2.26 mmol) and sodium hydrogencarbonate (4.10 g, 48.8 mmol) inacetone (125 mL) and water (45 mL) was added Oxone (3.47 g, 5.64 mmol).The reaction mixture was stirred at room temperature for 18 hours. Themixture was treated with excess sodium hydrosulfide, stirred for 15minutes, and extracted with ethyl acetate (3×50 mL). The combinedorganic phases were washed with brine (100 mL), dried (magnesiumsulfate), filtered, and concentrated to afford a white solid. The crudeproduct was triturated with hot methanol followed by flashchromatography over silica (methanol/methylene chloride) to provide0.230 g (23%) of the product as a white solid. ¹H NMR analysis wasconsistent with assigned structure.

Example 106-[4-(Chloro-benzylsulfanyl)-3,5-dimethyl-pyrazol-1-yl]-[2,2′]bipyridinyl(Compound 126) Step 1) Preparation of4-(4-Chloro-benzylsulfanyl)-3,5-dimethyl-1H-pyrazole

To a solution of crude 3-(4-chloro-benzylsulfanyl)-pentane-2,4-dione[prepared as described above from 3-chloro-pentan-2,3-dione (1.03 g,7.63 mmol) and 4-chlorobenzyl mercaptan (1.21 g, 7.63 mmol)] in ethanol(60 mL) was added hydrazine hydrate (0.713 mL, 22.9 mmol) andp-toluenesulfonic acid monohydrate (ca. 25 mg). The solution was heatedto reflux and stirred for 15 hours. The reaction mixture was allowed tocool to room temperature, and water (300 mL) was added. The precipitatethat formed was removed by filtration, washed with water, and dried toprovide 1.38 g (72%) of the product as a white solid: ¹H NMR (CDCl₃) δ7.17 (d, J=8.4 Hz, 2H), 6.92 (d, J=8.4 Hz, 2H), 3.57 (s, 2H), and 2.02(s, 6H) ppm. ¹³C NMR (CDCl₃) δ 137.2, 132.7, 130.3, 128.3, 105.2, 39.9,10.7 ppm.

Step 2) Preparation of Compound 126

To a solution of 4-(4-chloro-benzylsulfanyl)-3,5-dimethyl-1H-pyrazole(0.708 g, 2.80 mmol) in 2-methoxyethyl ether (7 mL) was added sodiumhydride (0.123 g (60% dispersion), 3.08 mmol). After stirring for 5minutes, 6-bromo-[2,2′]bipyridinyl (0.691 g, 2.94 mmol) was added, andthe reaction mixture was heated to 100° C. for 16 hours. The reactionmixture was allowed to cool to room temperature, and water (100 mL) wasadded. The suspension was extracted with ethyl acetate (50 mL×2), andthe combined organic extracts washed with brine (100 mL). The organicphase was dried (magnesium sulfate), filtered, and concentrated toafford a tan solid. Flash chromatography over silica (methanol/methylenechloride) followed by recrystallization from aqueous methanol provided0.485 g (43%) of the product as a white solid: ¹H NMR (CDCl₃) δ8.70-8.68 (m, 1H), 8.34 (d, J=7.9 Hz, 1H), 8.29 (d, J=7.9 Hz, 1H), 7.94(t, J=7.9 Hz, 1H), 7.86 (dt, J=2.0, 7.9 Hz, 2H), 7.35-7.31 (m, 1H), 7.22(d, J=8.3 Hz, 2H), 6.99 (d, J=8.3 Hz, 2H), 3.65 (s, 2H), 2.45 (s, 3H),2.21 (s, 3H) ppm. ¹³C NMR (CDCl₃) δ 155.7, 154.5, 153.8, 152.9, 149.5,145.9, 139.7, 137.3, 137.2, 133.1, 130.7, 128.7, 124.2, 121.3, 118.5,116.1, 110.0, 40.3, 13.6, 12.3 ppm.

Example 114-[5-Benzyl-3-(4-chloro-phenyl)-[1,2,4]triazol-1-yl]-6-pyridin-2-yl-[1,3,5]triazin-2-ylamine(Compound 133) Step 1) Preparation ofN-(4-Chloro-thiobenzoyl)-2-phenyl-acetamide

To a solution of 4-chlorothiobenzamide (0.519 g, 3.02 mmol) in acetone(5 mL) was added pyridine (0.367 mL, 4.53 mmol) and phenylacetylchloride (0.480 mL, 3.63 mmol). The bright orange reaction mixture washeated to 55° C. for 1 hour. The reaction mixture was allowed to cool toroom temperature, and water (20 mL) was added. The precipitate wasremoved by filtration, washed with water, and dried to provide 0.721 g(82%) of the product as a red solid: ¹H NMR (CDCl₃) δ 9.34 (br s, 1H),7.50-7.22 (m, 9H), 3.96 (s, 2H) ppm.

Step 2) Preparation of Compound 133

To a solution of N-(4-chloro-thiobenzoyl)-2-phenyl-acetamide (0.536 g,1.85 mmol) and sodium acetate (ca. 0.25 g) in 1:1 aceticacid/1,4-dioxane (30 mL) was added4-hydrazino-6-pyridin-2-yl-[1,3,5]triazin-2-ylamine (0.376 g, 1.85mmol). The solution was heated to 90° C. and stirred for 15 hours. Thereaction mixture was allowed to cool to room temperature, and water (100mL) was added. The precipitate was removed by filtration, trituratedwith hot ethanol (300 mL), and dried to provide 0.489 g (60%) of theproduct as a white solid: ¹H NMR (DMSO-d₆) δ 8.78 (d, J=4.3 Hz, 1H),8.32 (d, J=6.1 Hz, 2H), 8.22 (d, J=7.8 Hz, 1H), 8.06 (d, J=8.5 Hz, 2H),7.96 (dt, J=1.5, 7.8 Hz, 1H), 7.62-7.57 (m, 3H), 7.36-7.26 (m, 2H),7.24-7.22 (m, 2H), 7.19-7.16 (m, 1H), 4.90 (s, 2H) ppm.

Example 122-[4-(4-Chloro-phenylsulfanyl)-3,5-dimethyl-pyrazol-1-yl]-4-pyridin-2-yl-[1,3,5]triazine(Compound 141) Step 1) Preparation of4-Pyridin-2-yl-[1,3,5]triazine-2-thiol

To a stirred solution of picolamide (1.50 g, 12.3 mmol) inN,N-dimethylformamide (20 mL) was addedtert-butoxybis(dimethylamino)methane (5.2 mL, 25 mmol). The mixture wasrefluxed for 2.5 hours and then concentrated. The resulting viscousbrown oil was taken up in a 0.5 M solution of sodium ethoxide in ethanol(50 mL, 25 mmol). Thiourea (0.72 g, 9.5 mmol) was added and the solutionwas refluxed for 2.5 hours. The reaction was concentrated and theresidue partitioned between ethyl acetate and dilute aqueous sodiumhydroxide solution. The aqueous layer was washed twice with ethylacetate and neutralized with 1 N aqueous hydrochloric acid. Theresulting precipitate was filtered off and rinsed with water. Vacuumoven dying afforded 0.72 g (40%) of crude product as a fine brown solid.This material was used without further purification in the next step.

Step 2) Preparation of 2-Methylsulfanyl-4-pyridin-2-yl-[1,3,5]triazine

To a stirred suspension of the product of step 1 (0.707 g, 3.72 mmol) inacetone (36 mL) was added sodium carbonate (0.79 g, 7.5 mmol) followedby iodomethane (0.35 mL, 5.6 mmol). After overnight stirring, thereaction was filtered free of undissolved solid and concentrated toafford product as light brown solid: ¹H NMR (DMSO-d₆) δ 9.13 (s, 1H),8.84-8.76 (m, 1H), 8.55-8.43 (m, 1H), 8.11-8.00 (m, 1H), 7.69-7.60 (m,1H), 2.63 (s, 3H) ppm.

Step 3) Preparation of (4-Pyridin-2-yl-[1,3,5]triazin-2-yl)-hydrazine

A stirred solution of the product of step 2 (0.763 g, 3.74 mmol) andhydrazine hydrate (0.22 mL, 3.9 mmol) in ethanol (8 mL) was heated atreflux for 30 minutes. The reaction was cooled in an ice bath and theprecipitate was filtered off. Vacuum oven drying afforded crude productas light brown solid. This material was used without furtherpurification in the next step.

Step 4) Preparation of Compound 141

A stirred suspension of the product of step 3 (0.333 g, 1.77 mmol) and3-(4-chloro-phenylsulfanyl)-pentane-2,4-dione (0.473 g, 1.95 mmol) inn-propanol was heated at 95° C. overnight. The reaction solution wasconcentrated directly onto silica and flash chromatographed (methylenechloride/methanol) to afford a dirty yellow solid. This material wasfurther purified by trituration with diethyl ether to afford 0.056 g(8%) of product as a white solid: ¹H NMR (DMSO-d₆) δ 9.44 (s, 1H),8.94-8.77 (m, 1H), 8.68-8.43 (m, 1H), 8.18-7.96 (m, 1H), 7.77-7.57 (m,1H), 7.45-7.24 (m, 2H), 7.20-6.98 (m, 2H), 2.83 (s, 3H), 2.20 (s, 3H)ppm.

Example 134-(4-Benzyl-3,5-dimethyl-pyrazol-1-yl)-6-pyridin-2-yl-[1,3,5]triazin-2-ylamine

(Compound 142)

Step 1) Preparation of 3-Benzylpentane-2,4-dione

To a solution of sodium ethoxide (freshly prepared from sodium hydride(0.474 g, 19.8 mmol) and anhydrous ethanol (50 mL)) was addedpentane-2,4-dione (6.00 g, 60.0 mmol). The resulting mixture was heatedto 50° C. while a solution of benzyl bromide (3.42 g, 20 mmol) inethanol (20 mL) was added over 30 minutes. The reaction was then heatedto reflux. After 2 hours, the mixture was concentrated, and the residuewas dissolved in ethyl acetate, washed three times with water, once withbrine, dried (magnesium sulfate) and concentrated to afford an oil.Flash chromatography over silica (ethyl acetate/dichloromethane)afforded 3.13 g (82%) of a colorless oil. The proton spectrum showed theproduct exists as equal parts of the keto and enol tautomers: ¹H NMR(CDCl₃) δ 16.81 (s, 0.5H), 7.13-7.32 (m, 5H), 4.00 (t, J=7.6 Hz, 0.5H),3.65 (s, 1H), 3.14 (d, J=7.6 Hz, 0.5H), 2.06-2.13 (m, 6H) ppm.

Step 2) Preparation of Compound 142

A suspension of 4-hydrazino-6-pyridin-2-yl-[1,3,5]triazin-2-ylamine(5.25 g, 25.6 mmol), 3-benzylpentane-2,4-dione (5.0 g, 28.4 mmol), andp-toluenesulfonic acid monohydrate (0.475 g, 2.50 mmol) indimethylsulfoxide (50 mL) was heated to 75° C. for 20 hours. Thereaction mixture was allowed to cool to room temperature, and thesuspension was dissolved in methylene chloride (700 mL). The solutionwas washed three times with water, once with brine, dried (magnesiumsulfate), and concentrated to afford a moist solid. The crude productwas suspended in ethyl acetate (50 mL), heated to reflux, allowed tocool to room temperature, and filtered (repeated three times). The finalproduct was obtained as a white solid containing 3% dimethylsulfoxide byweight: 8.63 g (90%). ¹H NMR (CDCl₃) δ 8.84-8.53 (m, 1H), 8.48 (d, J=7.9Hz, 1H), 7.86-7.91 (m, 1H), 7.46-7.50 (m, 1H), 7.13-7.30 (m, 5H), 6.61(br s, 1H), 6.40 (br s, 1H), 3.83 (s, 2H), 2.79 (s, 3H), 2.24 (s, 3H)ppm. MS (ESI) m/z 358 (M+H⁺).

Example 144-[3,5-Dimethyl-4-(3-phenyl-propyl)-pyrazol-1-yl]-6-pyridin-2-yl-[1,3,5]triazin-2-lyamine(Compound 144) Step 1) Preparation of3-(3-Phenyl-propyl)-pentane-2,4-dione

A mixture of pentane-2,4-dione (3.87 g, 38.6 mmol),3-phenyl-1-iodopropane (3.18 g, 12.9 mmol) and anhydrous potassiumcarbonate (1.7 g, 12.3 mmol) in acetone (7.5 mL) was heated to refluxfor 24 hours. The room temperature reaction mixture was filtered, andthe filter cake washed with acetone (25 mL×3). The combined filtrateswere concentrated, and the residue was partitioned between ethyl acetateand water. The organic layer was washed with water and brine, dried(magnesium sulfate), and concentrated to an oil. Flash chromatographyover silica (ethyl acetate/hexanes) afforded 1.13 g (42%) of a colorlessoil. The proton spectrum in deuterochloroform showed the product existsas a mixture of keto and enol tautomers.

Step 2) Preparation of Compound 144

A solution of 4-hydrazino-6-pyridin-2-yl-[1,3,5]triazin-2-ylamine (6.17g, 30.4 mmol), 3-(3-phenyl-propyl)-pentane-2,4-dione (6.66 g, 30.4mmol), and p-toluenesulfonic acid monohydrate (0.05 g, 0.26 mmol) indimethylsulfoxide (200 mL) was heated 75° C. for 18 hours. The solutionwas allowed to cool to room temperature then diluted with ethyl acetate(700 mL). The resulting solution was washed 3 times with water, oncewith brine, dried (magnesium sulfate) and concentrated to afford a creamcolored solid. The solid was triturated in boiling ethyl acetate (200mL), and the suspension allowed to cool to room temperature. After 2hours, the product was collected by filtration and dried in vacuo toyield 5.87 g (50%) of a white solid: ¹H NMR (CDCl₃) δ 8.84-8.54 (m, 1H),8.49 (d, J=7.9 Hz, 1H), 7.87-7.91 (m, 1H), 7.46-7.50 (m, 1H), 7.18-7.32(m, 5H), 6.44 (br s, 1H), 6.25 (br s, 1H), 2.73 (s, 3H), 2.65-2.73 (m,2H), 2.44-2.49 (m, 2H), 2.31 (s, 3H), 1.79-1.86 (m, 2H) ppm. MS (ESI)m/z 386 (M+H⁺).

Example 154-(4-Chloro-phenylsulfanyl)-5-methyl-2-(4-pyridin-2-yl-pyrimidin-2-yl)-2,4-dihydro-pyrazol-3-one(Compound 147) Step 1) Preparation of2-(4-Chloro-phenylsulfanyl)-3-oxo-butyric acid ethyl ester

To a solution of ethyl 2-chloroacetoacetate (1.58 g, 9.61 mmol) inethylene glycol dimethyl ether (30 mL) was added sodiumhydrogencarbonate (ca. 15 g) and 4-chlorothiophenol (1.39 g, 9.61 mmol).The suspension was heated to reflux and stirred for 3 hours. Thereaction mixture was allowed to cool to room temperature, and water (100mL) was added. The mixture was extracted with ethyl ether (75 mL), andthe organic extract was washed with brine (150 mL). The organic phasewas dried (magnesium sulfate), filtered, and concentrated to afford ayellow oil. Flash chromatography over silica (ethyl acetate/hexanes)provided 1.44 g (55%) of the product as a colorless oil: ¹H NMR (CDCl₃)δ 13.8 (s, 1H), 7.26-7.21 (m, 2H), 7.06-7.04 (m, 2H), 4.21 (q, J=7.1 Hz,2H), 2.33 (s, 3H), 1.20 (t, J=7.1 Hz, 3H).

Step 2) Preparation of Compound 147

To a solution of 2-(4-chloro-phenylsulfanyl)-3-oxo-butyric acid ethylester (4.13 g, 15.1 mmol) in n-propanol (85 mL) was added(4-pyridin-2-yl-pyrimindin-2-yl)-hydrazine (2.84 g, 15.1 mmol) andp-toluenesulfonic acid monohydrate (ca. 25 mgs). The solution was heatedto reflux and stirred for 15 hours. The reaction mixture was allowed tocool to room temperature, diluted with water (400 mL), and extractedwith ethyl acetate (250 mL). The organic phase was washed water (200 mL)and brine (200 mL), dried (magnesium sulfate), filtered, andconcentrated to afford a brown oil. The crude material was trituratedwith diethyl ether, and the solid that formed was isolated byfiltration. Further trituration with a minimum amount of hot methanolprovided the product as a tan solid. ¹H NMR (CDCl₃) δ 12.8 (br s, 1H),8.85 (d, J=5.3 Hz, 1H), 8.77 (d, J=4.3 Hz, 1H), 8.44 (d, J=7.8 Hz, 1H),8.29 (d, J=5.3 Hz, 1H), 7.92 (dt, J=1.5, 7.8 Hz, 1H), 7.51-7.48 (m, 1H),7.19 (d, J=8.6 Hz, 2H), 7.09 (d, J=8.6 Hz, 2H), 2.32 (s, 3H). ¹³C NMR(CDCl₃) δ 165.1, 159.1, 158.8, 157.1, 156.8, 152.0, 137.7, 136.6, 131.3,129.2, 127.3, 126.8, 122.7, 114.6, 88.9, 13.3.

Example 162-[4-(4-Chloro-phenylsulfanyl)-5-methoxy-3-methyl-pyrazol-1-yl]-4-pyridin-2-yl-pyrimidine(Compound 148)

To a solution of4-(4-chloro-phenylsulfanyl)-5-methyl-2(4-pyridin-2yl-pyrimidin-2-yl)-2,4-dihydro-pyrazol-3-one(0.344 g, 0.869 mmol) in acetone (10 mL) was added potassium carbonate(ca. 3 g) and iodomethane (0.130 g, 0.912 mmol). The suspension washeated to reflux and stirred for 2 hours. The suspension was allowed tocool to room temperature, and water (100 mL) was added. The mixture wasextracted with diethyl ether (50 mL), and the organic phase was washedwith brine (50 mL), dried (magnesium sulfate), filtered and concentratedto afford a white solid. Flash chromatography over silica (methylenechloride/2M ammonia in methanol, two columns) provided 0.007 g (2%) ofthe product as a white solid. ¹H NMR analysis was consistent with theassigned structure.

TABLE I

Meth- Cpd od No. Used R₁₁ R₁₀ R₉ R₈ R₁₂  1 1

H  2 1

H  3 1

H  4 1

H  5 1

H  6 1

H  7 1

H  8 1

H  9 1

H 10 1

H 11 1

H 12 1

H 13 1

H 14 1

H 15 1

H 16 1

H 17 1

H 18 1

H 19 1

H 20 1

H 21 1

H 22 1

H 23 1

H 24 1

H 25 1

H 26 4

CH3 27 1

H 28 1

H 29 1

H 30 1

H 31 1

H 32 1

H 33 1

H 34 1

H 35 1

H 36 1

H 37 1

H 38 1

H 39 1

H 40 1

H 41 1

H 42 1

H 43 1

H 44 1

H 45a 3

H 46a 3

H 47b 3

H 48b 3

H 49c 1

H 50c 1

H 51 2

H 52 2

H 53 2

H 54 2

H 55 2

H 56 2

H 57 2

H 58 2

H 59 2

H,H

H 60 2

H 61 2

H 62 2

H 63 2

H 64 2

H 65 2

H 66 2

H 67 2

H 68 2

H 69 2

H 70 2

H 71 2

H 72 2

H 73 2

H 74 2

H 75 2

H 76 2

H 77 2

H 78 2

H 79 2

H 80 2

H 81 2

H 82 2

H 83 2

H 84 2

H 85 2

H 86 2

H 87 2

H 88 2

H 89 2

H 90 2

H 91 2

H 92 2

H aCompound 45 was prepared at the (R)-enantiomer. Compound 46 wasprepared as the (S)-enantiomer. Enantiomeric excesses were determined tobe 91.4% and 91.8% respectively. bCompound 47 was prepared at the(R)-enantiomer. Compound 48 was prepared as the (S)-enantiomer.Enantiomeric excesses were determined to be 100% (within detectionlimits) and >95% respectively. cCompounds 49 and 50 are the (−) and (+)enantiomers, respectively, of compound 44. Preparative chiral HPLC wasused for the resolution (ChiralPack OD, 2 × 25 cm; eluant: 40/60 carbondioxide/acetonitrile; detection 260 nM).

TABLE II

Cpd Ex. No. No. R₁₉ R₄ R₃ n X W V  93  6 H H CH₃ 0 O N CH  94  6 4-Cl HCH₃ 0 O N CH  95  6 4-F H CH₃ 0 O N CH  96  6 4-CH₃O H CH₃ 0 O N CH  97 6 4-(CH₃)₂CH H CH₃ 0 O N CH  98  6 4-CF₃ H CH₃ 0 O N CH  99  6 4-CF₃O HCH₃ 0 O N CH 100  7 4-(CH₂OH) H CH₃ 0 O N CH 101  7 4-CHO H CH₃ 0 O N CH102  7 4-(N-morpholino)CH₂ H CH₃ 0 O N CH 103  7 4-(N- H CH₃ 0 O N CHtetrahydroisoquinolino)CH₂ 104  7 4-(4- H CH₃ 0 O N CHBenzyl-piperazin-1-yl)CH₂ 105  7 4-(4-(2-fluoro-phenyl) H CH₃ 0 O N CHpiperazin-1-yl)CH₂ 106  6 4-Cl H CH₃ 0 O N N 107  6 4-Cl H CH₃ 1 O N CH108  8 H H CH₃ 0 S N CH 109  8 2-Cl H CH₃ 0 S N CH 110  8 3-Cl H CH₃ 0 SN CH 111  8 4-Cl H CH₃ 0 S N CH 112  8 4-F H CH₃ 0 S N CH 113  8 4-CH₃OH CH₃ 0 S N CH 114  8 3,4-diCl H CH₃ 0 S N CH 115  8 4-Cl 4-CH₃ CH₃ 0 SN CH 116  8 4-Cl 5-CH₃ CH₃ 0 S N CH 117  8 H H CH₃ 1 S N CH 118  8 2-ClH CH₃ 1 S N CH 119  8 4-Cl H CH₃ 1 S N CH 120  8 4-F H CH₃ 1 S N CH 121 8 4-CH₃ H CH₃ 1 S N CH 122  8 4-CH₃O H CH₃ 1 S N CH 123  8 4-CF₃O H CH₃1 S N CH 124  9 4-Cl H CH₃ 1 SO₂ N CH 125 10 4-Cl H CH₃ 0 O C CH 126 104-Cl H CH₃ 1 S C CH 147 15 4-Cl H OH 0 S N CH 148 16 4-Cl H OCH₃ 0 S NCH

TABLE III

Method Cmp No. Used R₂₁ R₂₂ 127 11 2,4-diClPh 4-ClPhCH₂ 128 11 2,4-diFPh4-ClPhCH₂ 129 11 4-t-BuPh 4-ClPhCH₂ 130 11 4-ClPh 4-ClPhCH₂ 131 112,4-diFPh 4-(CH₃O)PhCH₂ 132 11 4-ClPh 3-(CH₃O)PhCH₂ 133 11 4-ClPh PhCH₂134 11 4-ClPh c-C₆H₁₁ 135 11 4-ClPh c-(C₅H₉)CH₂CH₂ 136 11 4-ClPh c-C₅H₉137 11 4-ClPh (CH₃)₃CCH₂ 138 11 4-C1Ph 3,4-di(CH₃O)PhCH₂

TABLE IV

Method Cpd No. Used R₂₃ R₂₄ n X 139  8 4-Cl NH₂ 0 S 140  9 4-Cl NH₂ 0SO₂ 141 12 4-Cl H 0 S 142 13 H NH₂ 0 C 143 13 4-Cl NH₂ 1 C 144 14 H NH₂2 C 145 13 4-Cl NH₂ 1 C 146 14 H NH₂ 2 C

II. Biological Results Example 17 In Vitro Activity of Compounds inTNF-α and VCAM Assays

Description of Table V: The results shown in Table V were obtained usingthe TNF-α assays performed as described in the protocols entitled“Method for TNF High Throughput Screen” and “Method for VCAM HighThroughput Screen”, which are detailed below. These assays measure acellular response to TNF-α stimulation, and the data published in thetable are expressed as percentage inhibitions. Percentage inhibition iscomputed on a scale of 0 to 100% where 0 percent inhibition means thecompound has no effect in the assay (the response is indistinguishablefrom TNF-α treatment alone), and 100% inhibition means that when thecompound is present at the stated concentration, the response in theassay is the same as if TNF-α were not added. These two assays measuretwo different outcomes of TNF-α treatment, cell death and Vascular CellAdhesion Molecule (VCAM) expression, and capture two of TNF-α's manybiological effects. TNF-α induced apoptosis is a mechanism by whichTNF-α production during immune/inflammatory responses can lead to directcellular and tissue damage. TNF-α VCAM expression in endothelial cellsis a response that allows leukocytes to leave the blood and enter a siteof immune/inflammatory response. Both of these activities could lead topathology in immune/inflammatory diseases. In the first column of thetable, the percentage inhibition of the TNF-α driven apoptosis assaywhen the compounds are tested at 1 μM concentration is presented. Thesecond column in the table lists the IC50 for the compound, a standardmeasure using a dilution series in the assay, and fitting a sigmoidalcurve to the resulting dose-response curve. The IC50 is theconcentration (derived analytically from the sigmoidal function) atwhich the compound would inhibit 50% of the response in the assay. Thethird and fourth column list the corresponding analysis for the TNF-αdriven VCAM assay. In Table VI, data corresponding to the data in column1 of Table V is shown, with human cells replacing the mouse cells usedin the high throughput screen. It will be understood that the tabledemonstrates that a class of compounds can have its best activity in oneassay or the other, or that there can be crossover such that thecompounds can have activity to some degree in both assays. While notbeing bound by theory, we believe this data shows that the compounds canhave a modulating effect in more than one activity. As a consequence,the compounds will have use in treating more than one TNF-α mediatedcondition, and may be selected according to the manner in which thecondition manifests. This corresponds to the use of these compounds inthe treatment of human disease.

TNF High Throughput Screen.

Protocol:

-   1. Plate 4×10⁴ L929 cells in 100 μl of complete EMEM in Costar 96    well plates in PM.-   2. The next day, pretreat with compound, inhibitor and control    vehicle for 2 hours.-   3. After 2 hours pretreatment, add 10 μl 5 ng/ml human TNF-α and    actinomycin D (40 μg/ml final concentration).-   4. Incubate overnight.-   5. In AM, remove supernatant.-   6. Wash on a plate washer (0.9% NaCl).-   7. Add 100 μl Crystal Violet 0.1% in 20% ETOH.-   8. Incubate at RT for 10 minutes.-   9. Wash on plate washer.-   10. Air dry wells at 37° C.-   11. Add 100 μl of methanol to each well.-   12. Shake plates on orbital shaker and read at 595 nm on plate    reader.    VCAM High Throughput Screen:    Protocol:-   1. Plate Primary human umbilical vein endothelial cells at 1.8×10⁴    cells/well in a 96-well tissue culture plate.-   2. Return Plates to 37° C. incubator for 48-72 hours before assay.-   3. On assay day, aspirate wells and add 1804, of endothelial growth    cell medium (EGM) to each well.-   4. Add Compound to each well.-   5. Shake plates for 3 minutes.-   6. Incubate plate for 1 hour at 37° C.-   7. Add Tumor Necrosis Factor (TNF) at 1.0 ng/ml final concentration.-   8. Shake plates for 3 minutes.-   9. Incubate plate for 2 hours at 37° C.-   10. Remove plate from incubator and wash plate 3 times with    phosphate buffered saline (PBS) using a plate washer.-   11. Add anti-VCAM antibody at 0.5 μg/ml final concentration.-   12. Incubate at 4° C. overnight.-   13. Wash 3 times with PBS on plate washer.-   14. Add goat anti-mouse horseradish peroxidase conjugate.-   15. Incubate at room temperature for one hour.-   16. Wash 3 times with PBS on plate washer.-   17. Add TMB to each well for 15 minutes at room temperature.-   18. Stop reaction with 100 μL of 2 N sulfuric acid.-   19. Read absorbance at 450 nm.

TABLE V In Vitro Activity of Compounds in TNF-α and VCAM Assays TNFαTNFα VCAM VCAM Cpd. % Inh. IC₅₀ % Inh. IC₅₀ No. @ 1 μM (μM) @ 12.5 μM(μM) 1 2.64 2 5.65 3 1.29 4 0.941 5 0.872 6 1.06 7 4.01 8 1.62 9 0.49610 3.68 11 2.89 12 5.15 13 1.37 14 1.79 15 0.386 16 1.32 17 1.5 18 2.6819 0.457 20 0.299 21 0.913 22 3.85 23 2.58 24 1.03 25 1.58 26 3.36 272.72 28 5.27 29 3.72 30 2.36 31 3.35 32 2.33 33 2.35 34 1.54 35 2.64 362.15 37 2.36 38 2.79 39 1.32 40 1.74 41 6.49 42 0.779 16 43 2.98 44 0.4124.4 45 3.34 46 4.37 47 5.58 48 4.05 49 1.53 50 0.39 51 3.56 52 3.49 537.13 54 6.03 55 7.64 56 6.63 57 56 58 50 59 51 60 50 61 51 62 59 63 5564 63 65 52 66 51 67 52 68 57 69 61 70 58 71 57 72 57 73 76 74 55 75 6776 56 77 54 78 60 79 62 80 53 81 50 82 55 83 56 84 52 85 52 86 53 87 5088 53 89 56 90 51 91 71 92 52 93 1.21 94 0.029 95 0.2 96 0.046 97 0.39798 0.058 99 0.177 100 3.46 101 17.9 102 0.431 103 0.709 104 0.05 1050.089 106 0.96 107 47 108 2.28 109 0.63 110 6.34 111 0.132 112 0.309 1130.498 114 1.44 115 19.4 116 355 117 16 118 72 119 7.95 120 13.2 121 9.93122 11.9 123 7.1 124 13.4 125 0.023 126 37.9 2.52 127 2.2 128 20.6 8.67129 19.5 130 8.96 131 12.8 132 1.39 133 19.5 134 0.526 135 0.275 1361.44 137 9.67 138  54^(a) 139  62^(b) 140 1.77 141 0.0011 142 0.152 1436.06 144 0.015 145 2.5 146 0.197 147 0.734 148 32.1 ^(a)Inhibitionmeasured at 2 μM ^(b)Inhibition measured at 25 μM

TABLE VI Compound 44 can inhibit the TNF-α induced apoptosis in primaryhuman fibroblasts OD 595 nm Standard Deviation % Rescue DMSO 0.189 0.005N/A TNF + DMSO 0.091 0.006 N/A 2 μM Compound 44 0.109 0.008 18.4 4 μMCompound 44 0.147 0.033 57.1 8 μM Compound 44 0.198 0.002 109.2

Normal human dermal fibroblasts were seeded into 96-well plates at 3×10⁴cells/well. Cells were pretreated for 2 hours with 2 m/ml Actinomycin Dand either Compound 44 or DMSO as a vehicle control. Cells were thenexposed to 2 ng/ml TNF-α for 24 hours and stained with crystalviolet/ethanol. Crystal violet was solubilized with methanol andabsorbance at 595 nm was read on a microtiter plate reader.

Example 18 In Vivo Activity of Compounds in Sepsis and IBD Models andPharmacokinetic Parameters

Sepsis Model:

Sepsis was induced in the C57/BL mouse by the intravenous injection of20 ng of lipopolysaccharide/animal plus 20 mgs d-galactosamine/animal.Inhibition was measured as the prevention of mortality over a three dayperiod. Compounds were administered intraperitoneally in 10%cremophore/10% ethanol/80% normal saline one hour before induction.

Inflammatory Bowel Disease Model:

Groups of three male rats weighing 150+/−10 g and fasted for 24 hourswere used. Distal colitis was induced by intracolonic instillation of0.5 ml/rat DNBS (2,4-dinitrobenzene sulfonic acid, 60 mg/ml in ethanol30%) after which air (2 ml) was gently injected through the cannula toensure that the solution remains in the colon. A test compound wasadministered orally 24 and 2 hours before DNBS-instillation and thendaily for 5 days in a total of 7 doses. The animals were sacrificed 24hours after the final dose of test compound administration and eachcolon was removed and weighed.

TABLE VII In Vivo Activity of Compounds in Sepsis and IBD Models andPharmacokinetic Parameters Sepsis: Inflammatory Murine LPS-d- BowelDisease: Pharmacokinetic galactosamine Rat DNBS model* Parameters model# % inhibition of colonic % oral % inhibition weight gain t^(1/2)bioavailability 44 33 +/− 1 @ 200 44 +/− 8 @ 100 mg/kg 2.3 26 mg/kg (2exp.) (2 exp.) 119 33 @ 200 mg/kg 51% @ 100 mg/kg 2.3 Undetectable 94Not active below 18% @ 20 mg/kg 10.9 Undetectable 200 mg/kg 43 33 @ 10mg/kg 31% @ 50 mg/kg 3.6 <1

Example 19 In Vivo Activity of Compounds in Experimental AllergicEncephalitis (Murine Model of Multiple Sclerosis)

SJL mouse strain were immunized subcutaneously with proteolipid peptideamino acid residues 139-151 in Complete Freund's Adjuvant. Pertussistoxin was injected intravenously on day 1 and 3 post-induction. Weightloss occurred at day 7 post-immunization with paralysis ensuing betweendays 9 and 14 post-immunization.

TABLE VIII In Vivo Activity of Compounds in Experimental AllergicEncephalitis (Murine Model of Multiple Sclerosis) Ratio of activitydisease severity score % % Survival⁷ (ADSS)⁴ at last inhibition oftreated vs. Dosing day of dosing for of % survival Compound regimen¹treated vs. control disease⁵ of controls 44 15 mgs/kg/day/i.p.² 1.9/3.35 44.0% 75 vs. 50 Induction regimen from day 1-21 P⁶ = 0.06 75mgs/kg/day/i.p.  0.8/3.35 75.7% 100 vs. 50  from day 1-21 p = 0.001 4415 mgs/kg/day/i.p. 2.6/4.6 43.5% 70 vs. 11 Therapeutic from day 7-21 p =0.08 regimen 75 mgs/kg/day/i.p. 1.1/4.6 76.1% 100 vs. 11  from day 7-21p = 0.001 44 75 mgs/kg/day/i.p. 2.2/4.4 50% 89 vs. 20 Oral therapeuticregimen from day 7-28 p = 0.002 94 75 mgs/kg/day/i.p. 2.15/3.53 39.1% 80vs. 67 Induction Regimen from day 1-18 p⁶ = 0.045 ¹Compounds solublizedin 10% cremophore and 10% ethanol 80% normal saline. ²Compoundsadministered intraperitoneally. ³Compound administered orally. ⁴ADSS isactivity disease severity score where >1 is significant disease.Paralysis starts at the tail and progresses towards head of the mousewhere a limp tail is a 1 and a complete paralysis is a 5. Animals areeuthanized at greater than or equal to a score of 4. ⁵% inhibition ofADSS = (average test ADSS − average control ADSS/control) × 100. ⁶Pvalues calculated using Student's T Test. ⁷Survival indicates thoseanimals remaining post-euthanasia.

EQUIVALENTS

While this invention has been particularly shown and described withreferences to preferred embodiments thereof, it will be understood bythose skilled in the art that various changes in form and details may bemade therein without departing from the scope of the inventionencompassed by the appended claims.

What is claimed is:
 1. A method of treating TNF-α mediated inflammatorybowel disease in a patient in need of such treatment, comprisingadministering to the patient a therapeutically effective amount of acompound of Formula (II):

or a physiologically acceptable salt thereof, wherein, R5 is substitutedor unsubstituted aralkyl, substituted or unsubstituted cycloalkyl, orsubstituted or unsubstituted cycloalkylalkyl; R6 is —H or —NR13R14; R7is substituted or unsubstituted phenyl; and R13 and R14 are each,independently, —H, a substituted or unsubstituted alkyl, a substitutedor unsubstituted cycloalkyl, a substituted or unsubstituted aryl, asubstituted or unsubstituted aralkyl; or R13 and R14 together with thenitrogen to which they are attached are a heterocycloalkyl.